Triazolyl macrocyclic hepatitis c serine protease inhibitors

ABSTRACT

The present invention relates to compounds of Formula I or II, or pharmaceutically acceptable salts, esters or prodrugs thereof: 
     
       
         
         
             
             
         
       
     
     which inhibit serine protease activity, particularly the activity of hepatitis C virus (HCV) NS3-NS4A protease. Consequently, the compounds of the present invention interfere with the life cycle of the hepatitis C virus and are also useful as antiviral agents. The present invention further relates to pharmaceutical compositions comprising the aforementioned compounds for administration to a subject suffering from HCV infection. The invention also relates to methods of treating an HCV infection in a subject by administering to the subject a pharmaceutical composition comprising a compound of the present invention.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. provisional application60/______ (conversion of U.S. application Ser. No. 11/503,487) filedAug. 11^(th), 2006, the entire content of which is herein incorporatedby reference.

TECHNICAL FIELD

The present invention relates to triazolyl macrocyclic hepatitis C virus(HCV) protease inhibitors having antiviral activity against HCV anduseful in the treatment of HCV infections. More particularly, theinvention relates to triazolyl macrocyclic HCV protease inhibitorcompounds, compositions containing such compounds and methods for usingthe same, as well as processes for making such compounds.

BACKGROUND OF THE INVENTION

HCV is the principal cause of non-A, non-B hepatitis and is anincreasingly severe public health problem both in the developed anddeveloping world. It is estimated that the virus infects over 200million people worldwide, surpassing the number of individuals infectedwith the human immunodeficiency virus (HIV) by nearly five fold. HCVinfected patients, due to the high percentage of individuals inflictedwith chronic infections, are at an elevated risk of developing cirrhosisof the liver, subsequent hepatocellular carcinoma and terminal liverdisease. HCV is the most prevalent cause of hepatocellular cancer andcause of patients requiring liver transplantations in the western world.

There are considerable barriers to the development of anti-HCVtherapeutics, which include, but are not limited to, the persistence ofthe virus, the genetic diversity of the virus during replication in thehost, the high incident rate of the virus developing drug-resistantmutants, and the lack of reproducible infectious culture systems andsmall-animal models for HCV replication and pathogenesis. In a majorityof cases, given the mild course of the infection and the complex biologyof the liver, careful consideration must be given to antiviral drugs,which are likely to have significant side effects.

Only two approved therapies for HCV infection are currently available.The original treatment regimen generally involves a 3-12 month course ofintravenous interferon-α (IFN-α), while a new approved second-generationtreatment involves co-treatment with IFN-α and the general antiviralnucleoside mimics like ribavirin. Both of these treatments suffer frominterferon related side effects as well as low efficacy against HCVinfections. There exists a need for the development of effectiveantiviral agents for treatment of HCV infection due to the poortolerability and disappointing efficacy of existing therapies.

In a patient population where the majority of individuals arechronically infected and asymptomatic and the prognoses are unknown, aneffective drug would desirably possess significantly fewer side effectsthan the currently available treatments. The hepatitis C non-structuralprotein-3 (NS3) is a proteolytic enzyme required for processing of theviral polyprotein and consequently viral replication. Despite the hugenumber of viral variants associated with HCV infection, the active siteof the NS3 protease remains highly conserved thus making its inhibitionan attractive mode of intervention. Recent success in the treatment ofHIV with protease inhibitors supports the concept that the inhibition ofNS3 is a key target in the battle against HCV.

HCV is a flaviridae type RNA virus. The HCV genome is enveloped andcontains a single strand RNA molecule composed of circa 9600 base pairs.It encodes a polypeptide comprised of approximately 3010 amino acids.

The HCV polyprotein is processed by viral and host peptidase into 10discreet peptides which serve a variety of functions. There are threestructural proteins, C, E1 and E2. The P7 protein is of unknown functionand is comprised of a highly variable sequence. There are sixnon-structural proteins. NS2 is a zinc-dependent metalloproteinase thatfunctions in conjunction with a portion of the NS3 protein. NS3incorporates two catalytic functions (separate from its association withNS2): a serine protease at the N-terminal end, which requires NS4A as acofactor, and an ATP-ase-dependent helicase function at the carboxylterminus. NS4A is a tightly associated but non-covalent cofactor of theserine protease.

The NS3-NS4A protease is responsible for cleaving four sites on theviral polyprotein. The NS3-NS4A cleavage is autocatalytic, occurring incis. The remaining three hydrolyses, NS4A-NS4B, NS4B-NS5A and NS5A-NS5Ball occur in trans. NS3 is a serine protease which is structurallyclassified as a chymotrypsin-like protease. While the NS serine proteasepossesses proteolytic activity by itself, the HCV protease enzyme is notan efficient enzyme in terms of catalyzing polyprotein cleavage. It hasbeen shown that a central hydrophobic region of the NS4A protein isrequired for this enhancement. The complex formation of the NS3 proteinwith NS4A seems necessary to the processing events, enhancing theproteolytic efficacy at all of the sites.

A general strategy for the development of antiviral agents is toinactivate virally encoded enzymes, including NS3, that are essentialfor the replication of the virus. Current efforts directed toward thediscovery of NS3 protease inhibitors were reviewed by S. Tan, A. Pause,Y. Shi, N. Sonenberg, Hepatitis C Therapeutics: Current Status andEmerging Strategies, Nature Rev. Drug Discov., 1, 867-881 (2002). Otherpatent disclosures describing the synthesis of HCV protease inhibitorsare: WO 00/59929 (2000); WO 99/07733 (1999); WO 00/09543 (2000); WO99/50230 (1999); U.S. Pat. No. 5,861,297 (1999); and US2002/0037998(2002).

SUMMARY OF THE INVENTION

The present invention relates to triazolyl macrocyclic HCV proteasecompounds, and including pharmaceutically acceptable salts, esters, orprodrugs thereof which inhibit serine protease activity, particularlythe activity of hepatitis C virus (HCV) NS3-NS4A protease. Consequently,the compounds of the present invention interfere with the life cycle ofthe hepatitis C virus and are also useful as antiviral agents. Thepresent invention further relates to pharmaceutical compositionscomprising the aforementioned compounds for administration to a subjectsuffering from HCV infection. The present invention further featurespharmaceutical compositions comprising a compound of the presentinvention (or a pharmaceutically acceptable salt, ester or prodrugthereof) and another anti-HCV agent, such as interferon (e.g.,alpha-interferon, beta-interferon, consensus interferon, pegylatedinterferon, or albumin or other conjugated interferon), ribavirin,amantadine, another HCV protease inhibitor, or an HCV polymerase,helicase or internal ribosome entry site inhibitor. The invention alsorelates to methods of treating an HCV infection in a subject byadministering to the subject a pharmaceutical composition of the presentinvention.

In one embodiment of the present invention there are disclosed compoundsrepresented by Formulae I or II, or pharmaceutically acceptable salts,esters, or prodrugs thereof:

wherein

A is selected from the group consisting of —(C═O)—O-Ri, —(C═O)—R₂,—C(═O)—NH—R₂, and —S(O)₂—R₁, —S(O)₂NHR₂;

R₁ is selected from the group consisting of:

(i) aryl; substituted aryl; heteroaryl; substituted heteroaryl;

(ii) heterocycloalkyl or substituted heterocycloalkyl; and

(iii) —C₁-C₈ alkyl, —C₂-C₈ alkenyl, or —C₂-C₈ alkynyl each containing 0,1, 2, or 3 heteroatoms selected from O, S, or N; substituted —C₁-C₈alkyl, substituted —C₂-C₈ alkenyl, or substituted —C₂-C₈ alkynyl eachcontaining 0, 1, 2, or 3 heteroatoms selected from O, S or N; —C₃-C₁₂cycloalkyl, or substituted —C₃-C₁₂ cycloalkyl; —C₃-C₁₂ cycloalkenyl, orsubstituted —C₃-C₁₂ cycloalkenyl;

R₂ is independently selected from the group consisting of:

(i) hydrogen;

(ii) aryl; substituted aryl; heteroaryl; substituted heteroaryl;

(iii) heterocycloalkyl or substituted heterocycloalkyl; and

(iv) —C₁-C₈ alkyl, —C₂-C₈ alkenyl, or —C₂-C₈ alkynyl each containing 0,1, 2, or 3 heteroatoms selected from O, S, or N; substituted —C₁-C₈alkyl, substituted —C₂-C₈ alkenyl, or substituted —C₂-C₈ alkynyl eachcontaining 0, 1, 2, or 3 heteroatoms selected from O, S or N; —C₃-C₁₂cycloalkyl, or substituted —C₃-C₁₂ cycloalkyl; —C₃-C₁₂ cycloalkenyl, orsubstituted —C₃-C₁₂ cycloalkenyl;

G is selected from the group consisting of —NHS(O)₂—R₃ and—NH(SO₂)NR₄R₅;

R₃ is selected from:

(i) aryl; substituted aryl; heteroaryl; substituted heteroaryl

(ii) heterocycloalkyl or substituted heterocycloalkyl;

(iii) —C₁-C₈ alkyl, —C₂-C₈ alkenyl, or —C₂-C₈ alkynyl containing 0, 1,2, or 3 heteroatoms selected from O, S or N, substituted —C₁-C₈ alkyl,substituted —C₂-C₈ alkenyl, or substituted —C₂-C₈ alkynyl containing 0,1, 2, or 3 heteroatoms selected from O, S or N; —C₃-C₁₂ cycloalkyl, orsubstituted —C₃-C₁₂ cycloalkyl; —C₃-C₁₂ cycloalkenyl, or substituted—C₃-C₁₂ cycloalkenyl;

provided that R₃ is not CH₂Ph or CH₂CH₂Ph;

R₄ and R₅ are independently selected from:

(i) hydrogen;

(ii) aryl; substituted aryl; heteroaryl; substituted heteroaryl;

(iii) heterocycloalkyl or substituted heterocycloalkyl; and

(iv) —C₁-C₈ alkyl, —C₂-C₈ alkenyl, or —C₂-C₈ alkynyl each containing 0,1, 2, or 3 heteroatoms selected from O, S, or N; substituted —C₁-C₈alkyl, substituted —C₂-C₈ alkenyl, or substituted —C₂-C₈ alkynyl eachcontaining 0, 1, 2, or 3 heteroatoms selected from O, S or N; —C₃-C₁₂cycloalkyl, or substituted —C₃-C₁₂ cycloalkyl; —C₃-C₁₂ cycloalkenyl, orsubstituted —C₃-C₁₂ cycloalkenyl;

L is selected from —CH₂—, —O—, —S—, and —S(O)₂—;

X and Y are independently selected from:

(i) hydrogen;

(ii) aryl; substituted aryl; heteroaryl; substituted heteroaryl;

(iii) heterocycloalkyl or substituted heterocycloalkyl;

(iv) —C₁-C₈ alkyl, —C₂-C₈ alkenyl, or —C₂-C₈ alkynyl each containing 0,1, 2, or 3 heteroatoms selected from O, S, or N; substituted —C₁-C₈alkyl, substituted —C₂-C₈ alkenyl, or substituted —C₂-C₈ alkynyl eachcontaining 0, 1, 2, or 3 heteroatoms selected from O, S or N; —C₃-C₁₂cycloalkyl, or substituted —C₃-C₁₂ cycloalkyl; —C₃-C₁₂ cycloalkenyl, orsubstituted —C₃-C₁₂ cycloalkenyl; and

(v) —W—R₆, where W is absent, or selected from —O—, —S—, —NH—, —N(Me)—,—C(O)NH—, and —C(O)N(Me)—; R₆ is selected from the group consisting of:

-   -   (a) hydrogen;    -   (b) aryl; substituted aryl; heteroaryl; substituted heteroaryl    -   (c) heterocycloalkyl or substituted heterocycloalkyl; and    -   (d) —C₁-C₈ alkyl, —C₂-C₈ alkenyl, or —C₂-C₈ alkynyl each        containing 0, 1, 2, or 3 heteroatoms selected from O, S or N;        substituted —C₁-C₈ alkyl, substituted —C₂-C₈ alkenyl, or        substituted —C₂-C₈ alkynyl each containing 0, 1, 2, or 3        heteroatoms selected from O, S or N; —C₃-C₁₂ cycloalkyl, or        substituted —C₃-C₁₂ cycloalkyl; —C₃-C₁₂ cycloalkenyl, or        substituted —C₃-C₁₂ cycloalkenyl;

alternatively, X and Y taken together with the carbon atoms to whichthey are attached to form a cyclic moiety which is selected from thegroup consisting of aryl, substituted aryl, heteroaryl, and substitutedheteroaryl;

denotes a carbon-carbon single or double bond;

j=0, 1, 2, 3,or 4;

k=1, 2, or 3;

m=0, 1, or 2; and

n=1,2or3.

DETAILED DESCRIPTION OF THE INVENTION

A first embodiment of the invention is a compound represented byFormulae I and II as described above, or a pharmaceutically acceptablesalt, ester or prodrug thereof, alone or in combination with apharmaceutically acceptable carrier or excipient.

In one embodiment of the invention is a compound represented by FormulaIII:

or a pharmaceutically acceptable salt, ester or prodrug thereof, aloneor in combination with a pharmaceutically acceptable carrier orexcipient, where A, Y, X and G are as defined in the previousembodiment.

In one example, X and Y are independently selected from the groupconsisting of hydrogen, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic, substituted heterocyclic, —C₁-C₈ alkyl, —C₂-C₈alkenyl, or —C₂-C₈ alkynyl, substituted —C₁-C₈ alkyl, substituted —C₂-C₈alkenyl, or substituted —C₂-C₈ alkynyl, —C₃-C₁₂ cycloalkyl, —C₃-C₁₂cycloalkenyl, substituted —C₃-C₁₂ cycloalkyl, and substituted —C₃-C₁₂cycloalkenyl, where each —C₁-C₈ alkyl, —C₂-C₈ alkenyl, —C₂-C₈ alkynyl,substituted —C₁-C₈ alkyl, substituted —C₂-C₈ alkenyl, and substituted—C₂-C₈ alkynyl independently contains 0, 1, 2, or 3 heteroatoms selectedfrom O, S, or N. A is selected from the group consisting of —C(O)—R₁,—C(O)—O—R₁ and —C(O)—NH—R₁, where R₁ is selected from aryl, substitutedaryl, heteroaryl, substituted heteroaryl, heterocyclic, substitutedheterocyclic, —C₁-C₈ alkyl, —C₂-C₈ alkenyl, —C₂-C₈ alkynyl, substituted—C₁-C₈ alkyl, substituted —C₂-C₈ alkenyl, substituted —C₂-C₈ alkynyl,—C₃-C₁₂ cycloalkyl, —C₃-C₁₂ cycloalkenyl, substituted —C₃-C₁₂cycloalkyl, or substituted —C₃-C₁₂ cycloalkenyl. G can be —NH—SO₂—NR₄R₅or —NHSO₂—R₃, where R₃ is selected from —C₁-C₈ alkyl, —C₂-C₈ alkenyl,—C₂-C₈ alkynyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic, substituted heterocyclic, —C₃-C₁₂ cycloalkyl,—C₃-C₁₂ cycloalkenyl, substituted —C₃-C₁₂ cycloalkyl, or substituted—C₃-C₁₂ cycloalkenyl, and R₄ and R₅ are each independently selected fromhydrogen, —C₁-C₈ alkyl, —C₂-C₈ alkenyl, —C₂-C₈ alkynyl, substituted—C₁-C₈ alkyl, substituted —C₂-C₈ alkenyl, substituted —C₂-C₈ alkynyl,aryl, substituted aryl, heteroaryl, substituted heteroaryl,heterocyclic, substituted heterocyclic, —C₃-C₁₂ cycloalkyl, —C₃-C₁₂cycloalkenyl, substituted —C₃-C₁₂ cycloalkyl, or substituted —C₃-C₁₂cycloalkenyl.

In still another example, X and Y are independently selected from thegroup consisting of hydrogen, aryl, substituted aryl, heteroaryl, andsubstituted heteroaryl. A is —C(O)—O—R₁ or —C(O)—NH—R₁, where R₁ is—C₁-C₈ alkyl, —C₂-C₈ alkenyl, —C₂-C₈ alkynyl, substituted —C₁-C₈ alkyl,substituted —C₂-C₈ alkenyl, substituted —C₂-C₈ alkynyl, —C₃-C₁₂cycloalkyl, —C₃-C₁₂ cycloalkenyl, substituted —C₃-C₁₂ cycloalkyl, orsubstituted —C₃-C₁₂ cycloalkenyl. G is —NHSO₂-R₃, where R₃ is selectedfrom aryl, substituted aryl, heteroaryl, substituted heteroaryl,heterocyclic, substituted heterocyclic, —C₃-C₁₂ cycloalkyl, —C₃-C₁₂cycloalkenyl, substituted —C₃-C₁₂ cycloalkyl, or substituted —C₃-C₁₂cycloalkenyl.

In still yet another example, X and Y are independently selected fromthe group consisting of aryl, substituted aryl, heteroaryl, andsubstituted heteroaryl. A is —C(O)—O—R₁, where R₁ is —C₃-C₁₂ cycloalkylor substituted —C₃-C₁₂ cycloalkyl. G is —NHSO₂—R₃, where R₃ is selectedfrom —C₃-C₁₂ cycloalkyl or substituted —C₃-C₁₂ cycloalkyl.

In another example, X and Y are independently selected from the groupconsisting of aryl, substituted aryl, heteroaryl, and substitutedheteroaryl. A is —C(O)—NH—R₁, where R₁ is —C₁-C₈ alkyl or substituted—C₁-C₈ alkyl. G is —NHSO₂-R₃, where R₃ is selected from —C₃-C₁₂cycloalkyl or substituted —C₃-C₁₂ cycloalkyl.

In still another example, X is substituted or unsubstituted aryl

Y is substituted or unsubstituted heteroaryl

A is selected from the group consisting of —C(O)—R₁, —C(O)—O—R₁ and—C(O)—NH—R₁, where R₁ is selected from aryl, substituted aryl,heteroaryl, substituted heteroaryl, heterocyclic, substitutedheterocyclic, —C₁-C₈ alkyl, —C₂-C₈ alkenyl, —C₂-C₈ alkynyl, substituted—C₁-C₈ alkyl, substituted —C₂-C₈ alkenyl, substituted —C₂-C₈ alkynyl,—C₃-C₁₂ cycloalkyl, —C₃-C₁₂ cycloalkenyl, substituted —C₃-C₁₂cycloalkyl, or substituted —C₃-C₁₂ cycloalkenyl. G can be —NH—SO₂—NR₄R₅or —NHSO₂—R₃, where R₃ is selected from —C₁-C₈ alkyl, —C₂-C₈ alkenyl,—C₂-C₈ alkynyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic, substituted heterocyclic, —C₃-C₁₂ cycloalkyl,—C₃-C₁₂ cycloalkenyl, substituted —C₃-C₁₂ cycloalkyl, or substituted—C₃-C₁₂ cycloalkenyl, and R₄ and R₅ are each independently selected fromhydrogen, —C₁-C₈ alkyl, —C₂-C₈ alkenyl, —C₂-C₈ alkynyl, substituted—C₁-C₈ alkyl, substituted —C₂-C₈ alkenyl, substituted —C₂-C₈ alkynyl,aryl, substituted aryl, heteroaryl, substituted heteroaryl,heterocyclic, substituted heterocyclic, —C₃-C₁₂ cycloalkyl, —C₃-C₁₂cycloalkenyl, substituted —C₃-C₁₂ cycloalkyl, or substituted —C₃-C₁₂cycloalkenyl.

In yet another example, X is substituted or unsubstituted aryl

Y is substituted or unsubstituted heteroaryl

A is —C(O)—O—R₁ or —C(O)—NH—R₁, where R₁ is —C₃—C₁₂ cycloalkyl orsubstituted —C₃-C₁₂ cycloalkyl. G is —NHSO₂—R₃, where R₃ is selectedfrom —C₃-C₁₂ cycloalkyl or substituted —C₃-C₁₂ cycloalkyl.

In another embodiment of the invention is a compound represented byFormula IV:

or a pharmaceutically acceptable salt, ester or prodrug thereof, aloneor in combination with a pharmaceutically acceptable carrier orexcipient, where A, Y, X and G are as defined in the first embodiment.

In one example, X and Y are independently selected from the groupconsisting of hydrogen, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic, substituted heterocyclic, —C₁-C₈ alkyl, —C₂-C₈alkenyl, or —C₂-C₈ alkynyl, substituted —C₁-C₈ alkyl, substituted —C₂-C₈alkenyl, or substituted —C₂-C₈ alkyny, —C₃-C₁₂ cycloalkyl, —C₃-C₁₂cycloalkenyl, substituted —C₃-C₁₂ cycloalkyl, and substituted —C₃-C₁₂cycloalkenyl, where each —C₁-C₈ alkyl, —C₂-C₈ alkenyl, —C₂-C₈ alkynyl,substituted —C₁-C₈ alkyl, substituted —C₂-C₈ alkenyl, and substituted—C₂-C₈ alkynyl independently contains 0, 1, 2, or 3 heteroatoms selectedfrom O, S, or N. A is selected from the group consisting of —C(O)—R₁,—C(O)—O—R₁ and —C(O)—NH—R₁, where R₁ is selected from aryl, substitutedaryl, heteroaryl, substituted heteroaryl, heterocyclic, substitutedheterocyclic, —C₁-C₈ alkyl, —C₂-C₈ alkenyl, —C₂-C₈ alkynyl, substituted—C₁-C₈ alkyl, substituted —C₂-C₈ alkenyl, substituted —C₂-C₈ alkynyl,—C₃-C₁₂ cycloalkyl, —C₃-C₁₂ cycloalkenyl, substituted —C₃-C₁₂cycloalkyl, or substituted —C₃-C₁₂ cycloalkenyl. G can be—NH—SO₂—NR₄R₅or —NHSO₂—R₃, where R₃ is selected from —C₁-C₈ alkyl,—C₂-C₈ alkenyl, —C₂-C₈ alkynyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, heterocyclic, substituted heterocyclic, —C₃-C₁₂cycloalkyl, —C₃-C₁₂ cycloalkenyl, substituted —C₃-C₁₂ cycloalkyl, orsubstituted —C₃-C₁₂ cycloalkenyl, and R₄ and R₅ are each independentlyselected from hydrogen, —C₁-C₈ alkyl, —C₂-C₈ alkenyl, —C₂-C₈ alkynyl,substituted —C₁-C₈ alkyl, substituted —C₂-C₈ alkenyl, substituted —C₂-C₈alkynyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl,heterocyclic, substituted heterocyclic, —C₃-C₁₂ cycloalkyl, —C₃-C₁₂cycloalkenyl, substituted —C₃-C₁₂ cycloalkyl, or substituted —C₃-C₁₂cycloalkenyl.

In still another example, X and Y are independently selected from thegroup consisting of hydrogen, aryl, substituted aryl, heteroaryl, andsubstituted heteroaryl. A is —C(O)—O—R₁ or —C(O)—NH—R₁, where R₁ is—C₁-C₈ alkyl, —C₂-C₈ alkenyl, —C₂-C₈ alkynyl, substituted —C₁-C₈ alkyl,substituted —C₂-C₈ alkenyl, substituted —C₂-C₈ alkynyl, —C₃-C₁₂cycloalkyl, —C₃-C₁₂ cycloalkenyl, substituted —C₃-C₁₂ cycloalkyl, orsubstituted —C₃-C₁₂ cycloalkenyl. G is —NHSO₂—R₃, where R₃ is selectedfrom aryl, substituted aryl, heteroaryl, substituted heteroaryl,heterocyclic, substituted heterocyclic, —C₃-C₁₂ cycloalkyl, —C₃-C₁₂cycloalkenyl, substituted —C₃-C₁₂ cycloalkyl, or substituted —C₃-C₁₂cycloalkenyl.

In still yet another example, X and Y are independently selected fromthe group consisting of aryl, substituted aryl, heteroaryl, andsubstituted heteroaryl. A is —C(O)—O—R₁, where R₁ is —C₃-C₁₂ cycloalkylor substituted —C₃-C₁₂ cycloalkyl. G is —NHSO₂—R₃, where R₃ is selectedfrom —C₃-C₁₂ cycloalkyl or substituted —C₃-C₁₂ cycloalkyl.

In another example, X and Y are independently selected from the groupconsisting of aryl, substituted aryl, heteroaryl, and substitutedheteroaryl. A is —C(O)—NH—R₁, where R₁ is —C₁-C₈ alkyl or substituted—C₁-C₈ alkyl. G is —NHSO₂—R₃, where R₃ is selected from —C₃-C₁₂cycloalkyl or substituted —C₃-C₁₂ cycloalkyl.

In one embodiment of the invention is a compound represented by FormulaV:

or a pharmaceutically acceptable salt, ester or prodrug thereof, aloneor in combination with a pharmaceutically acceptable carrier orexcipient, where X₁-X₄ are independently selected from —CR₇ and N,wherein R₇ is independently selected at each occurrence from:

-   -   (i) hydrogen; halogen; —NO₂; —CN;    -   (ii) —M—R₄, M is O, S, NH;    -   (iii) NR₄R₅;    -   (iv) —C₁-C₈ alkyl, —C₂-C₈ alkenyl, or —C₂-C₈ alkynyl each        containing 0, 1, 2, or 3 heteroatoms selected from O, S, or N;        substituted —C₁-C₈ alkyl, substituted —C₂-C₈ alkenyl, or        substituted —C₂-C₈ alkynyl each containing 0, 1, 2, or 3        heteroatoms selected from O, S or N; —C₃-C₁₂ cycloalkyl, or        substituted —C₃-C₁₂ cycloalkyl; —C₃-C₁₂ cycloalkenyl, or        substituted —C₃-C₁₂ cycloalkenyl;    -   (v) aryl; substituted aryl; heteroaryl; substituted heteroaryl;        and    -   (vi) heterocycloalkyl or substituted heterocycloalkyl;    -   A, G, R₄ and R₅ are are as defined in the first embodiment.

In one example, where X₁-X₄ are independently selected from —CR₇ and N,where R₇ is as previously defined immediately above. A is selected fromthe group consisting of —C(O)—R₁, —C(O)—O—R₁ and —C(O)—NH—R₁, where R₁is selected from aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic, substituted heterocyclic, —C₁-C₈ alkyl, —C₂-C₈alkenyl, —C₂-C₈ alkynyl, substituted —C₁-C₈ alkyl, substituted —C₂-C₈alkenyl, substituted —C₂-C₈ alkynyl, —C₃-C₁₂ cycloalkyl, —C₃-C₁₂cycloalkenyl, substituted —C₃-C₁₂ cycloalkyl, or substituted —C₃-C₁₂cycloalkenyl. G can be —NH—SO₂—NR₄R₅or —NHSO₂—R₃, where R₃ is selectedfrom —C₁-C₈ alkyl, —C₂-C₈ alkenyl, —C₂-C₈ alkynyl, aryl, substitutedaryl, heteroaryl, substituted heteroaryl, heterocyclic, substitutedheterocyclic, —C₃-C₁₂ cycloalkyl, —C₃-C₁₂ cycloalkenyl, substituted—C₃-C₁₂ cycloalkyl, or substituted —C₃-C₁₂ cycloalkenyl, and R₄ and R₅are each independently selected from hydrogen, —C₁-C₈ alkyl, —C₂-C₈alkenyl, —C₂-C₈ alkynyl, substituted —C₁-C₈ alkyl, substituted —C₂-C₈alkenyl, substituted —C₂-C₈ alkynyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, heterocyclic, substituted heterocyclic, —C₃-C₁₂cycloalkyl, —C₃-C₁₂ cycloalkenyl, substituted —C₃-C₁₂ cycloalkyl, orsubstituted —C₃-C₁₂ cycloalkenyl.

In still another example, where X₁-X₄ are independently selected from—CR₇ and N, where R₇ is as previously defined above. A is —C(O)—O—R₁ or—C(O)—NH—R₁, where R₁ is —C₁-C₈ alkyl, —C₂-C₈ alkenyl, —C₂-C₈ alkynyl,substituted —C₁-C₈ alkyl, substituted —C₂-C₈ alkenyl, substituted —C₂-C₈alkynyl, —C₃-C₁₂ cycloalkyl, —C₃-C₁₂ cycloalkenyl, substituted —C₃-C₁₂cycloalkyl, or substituted —C₃-C₁₂ cycloalkenyl. G is —NHSO₂—R₃, whereR₃ is selected from aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic, substituted heterocyclic, —C₃-C₁₂ cycloalkyl,—C₃-C₁₂ cycloalkenyl, substituted —C₃-C₁₂ cycloalkyl, or substituted—C₃-C₁₂ cycloalkenyl.

In still yet another example, where X₁-X₄ are independently selectedfrom —CR₇ and N, where R₇ is as previously defined above. A is—C(O)—O—R₁, where R₁ is —C₃-C₁₂ cycloalkyl or substituted —C₃-C₁₂cycloalkyl. G is —NHSO₂—R₃, where R₃ is selected from —C₃-C₁₂ cycloalkylor substituted —C₃-C₁₂ cycloalkyl.

In another example, where X₁-X₄ are independently selected from —CR₇ andN, where R₇ is as previously defined above. A is —C(O)—NH—R₁, where R₁is —C₁-C₈ alkyl or substituted —C₁-C₈ alkyl. G is —NHSO₂—R₃, where R₃ isselected from —C₃-C₁₂ cycloalkyl or substituted —C₃-C₁₂ cycloalkyl.

In one embodiment of the invention is a compound represented by FormulaVI:

or a pharmaceutically acceptable salt, ester or prodrug thereof, aloneor in combination with a pharmaceutically acceptable carrier orexcipient, where X₁-X₄ are independently selected from —CR₇ and N,wherein R₇ is independently selected at each occurrence from:

-   -   (i) hydrogen; halogen; —NO₂; —CN;    -   (ii) —M—R₄, M is O, S, NH;    -   (iii) NR₄R₅;    -   (iv) —C₁-C₈ alkyl, —C₂-C₈ alkenyl, or —C₂-C₈ alkynyl each        containing 0, 1, 2, or 3 heteroatoms selected from O, S, or N;        substituted —C₁-C₈ alkyl, substituted —C₂-C₈ alkenyl, or        substituted —C₂-C₈ alkynyl each containing 0, 1, 2, or 3        heteroatoms selected from O, S or N; —C₃-C₁₂ cycloalkyl, or        substituted —C₃-C₁₂ cycloalkyl; —C₃-C₁₂ cycloalkenyl, or        substituted —C₃-C₁₂ cycloalkenyl;    -   (v) aryl; substituted aryl; heteroaryl; substituted heteroaryl;        and    -   (vi) heterocycloalkyl or substituted heterocycloalkyl;    -   A, G, R₄ and R₅ are are as defined in the first embodiment.

In one example, where X₁-X₄ are independently selected from —CR₇ and N,where R₇ is as defined immediately above. A is selected from the groupconsisting of —C(O)—R₁, —C(O)—O—R₁ and —C(O)—NH—R₁, where R₁ is selectedfrom aryl, substituted aryl, heteroaryl, substituted heteroaryl,heterocyclic, substituted heterocyclic, —C₁-C₈ alkyl, —C₂-C₈ alkenyl,—C₂-C₈ alkynyl, substituted —C₁-C₈ alkyl, substituted —C₂-C₈ alkenyl,substituted —C₂-C₈ alkynyl, —C₃-C₁₂ cycloalkyl, —C₃-C₁₂ cycloalkenyl,substituted —C₃-C₁₂ cycloalkyl, or substituted —C₃-C₁₂ cycloalkenyl. Gcan be —NH—SO₂—NR₄R₅ or —NHSO₂—R₃, where R₃ is selected from —C₁-C₈alkyl, —C₂-C₈ alkenyl, —C₂-C₈ alkynyl, aryl, substituted aryl,heteroaryl, substituted heteroaryl, heterocyclic, substitutedheterocyclic, —C₃-C₁₂ cycloalkyl, —C₃-C₁₂ cycloalkenyl, substituted—C₃-C₁₂ cycloalkyl, or substituted —C₃-C₁₂ cycloalkenyl, and R₄ and R₅are each independently selected from hydrogen, —C₁-C₈ alkyl, —C₂-C₈alkenyl, -C₂-C₈ alkynyl, substituted —C₁-C₈ alkyl, substituted —C₂-C₈alkenyl, substituted —C₂-C₈ alkynyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, heterocyclic, substituted heterocyclic, —C₃-C₁₂cycloalkyl, —C₃-C₁₂ cycloalkenyl, substituted —C₃-C₁₂ cycloalkyl, orsubstituted —C₃-C₁₂ cycloalkenyl.

In still another example, where X₁-X₄ are independently selected from—CR₇ and N, where R₇ is as previously defined above. A is —C(O)—O—R₁ or—C(O)—NH—R₁, where R₁ is —C₁-C₈ alkyl, —C₂-C₈ alkenyl, —C₂-C₈ alkynyl,substituted —C₁-C₈ alkyl, substituted —C₂-C₈ alkenyl, substituted —C₂-C₈alkynyl, —C₃-C₁₂ cycloalkyl, —C₃-C₁₂ cycloalkenyl, substituted —C₃-C₁₂cycloalkyl, or substituted —C₃-C₁₂ cycloalkenyl. G is —NHSO₂—R₃, whereR₃ is selected from aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic, substituted heterocyclic, —C₃-C₁₂ cycloalkyl,—C₃-C₁₂ cycloalkenyl, substituted —C₃-C₁₂ cycloalkyl, or substituted—C₃-C₁₂ cycloalkenyl.

In still yet another example, where X₁-X₄ are independently selectedfrom —CR₇ and N, where R₇ is as previously defined above. A is—C(O)—O—R₁, where R₁ is —C₃-C₁₂ cycloalkyl or substituted —C₃-C₁₂cycloalkyl. G is —NHSO₂—R₃, where R₃ is selected from —C₃-C₁₂ cycloalkylor substituted —C₃-C₁₂ cycloalkyl.

In another example, where X₁-X₄ are independently selected from —CR₇ andN, where R₇ is as previously defined above. A is —C(O)—NH—R₁, where R₁is —C₁-C₈ alkyl or substituted —C₁-C₈ alkyl. G is —NHSO₂—R₃, where R₃ isselected from —C₃-C₁₂ cycloalkyl or substituted —C₃-C₁₂ cycloalkyl.

In one embodiment of the invention is a compound represented by FormulaVII:

or a pharmaceutically acceptable salt, ester or prodrug thereof, aloneor in combination with a pharmaceutically acceptable carrier orexcipient, where Y₁-Y₃ are independently selected selected from CR₇, N,NR₇, S and O, wherein R₇ is independently selected at each occurrencefrom:

-   -   (i) hydrogen; halogen; —NO₂; —CN;    -   (ii) —M—R₄, M is O, S, NH;    -   (iii) NR₄R₅;    -   (iv) —C₁-C₈ alkyl, —C₂-C₈ alkenyl, or —C₂-C₈ alkynyl each        containing 0, 1, 2, or 3 heteroatoms selected from O, S, or N;        substituted —C₁-C₈ alkyl, substituted —C₂-C₈ alkenyl, or        substituted —C₂-C₈ alkynyl each containing 0, 1, 2, or 3        heteroatoms selected from O, S or N; —C₃-C₁₂ cycloalkyl, or        substituted —C₃-C₁₂ cycloalkyl; —C₃-C₁₂ cycloalkenyl, or        substituted —C₃-C₁₂ cycloalkenyl;    -   (v) aryl; substituted aryl; heteroaryl; substituted heteroaryl;        and    -   (vi) heterocycloalkyl or substituted heterocycloalkyl;    -   A, G, R₄ and R₅ are are as defined in the first embodiment.

In one example, where Y₁-Y₃ are independently selected from —CR₇, N,NR₇, S and O, where R₇ is as previously defined immediately above. A isselected from the group consisting of —C(O)—R₁, —C(O)—O—R₁ and—C(O)—NH—R₁, where R₁ is selected from aryl, substituted aryl,heteroaryl, substituted heteroaryl, heterocyclic, substitutedheterocyclic, —C₁-C₈ alkyl, —C₂-C₈ alkenyl, —C₂-C₈ alkynyl, substituted—C₁-C₈ alkyl, substituted —C₂-C₈ alkenyl, substituted —C₂-C₈ alkynyl,—C₃-C₁₂ cycloalkyl, —C₃-C₁₂ cycloalkenyl, substituted —C₃-C₁₂cycloalkyl, or substituted —C₃-C₁₂ cycloalkenyl. G can be —NH—SO₂—NR₄R₅or —NHSO₂—R₃, where R₃ is selected from —C₁-C₈ alkyl, —C₂alkenyl, —C₂-C₈alkynyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl,heterocyclic, substituted heterocyclic, —C₃-C₁₂ cycloalkyl, —C₃-C₁₂cycloalkenyl, substituted —C₃-C₁₂ cycloalkyl, or substituted —C₃-C₁₂cycloalkenyl, and R₄ and R₅ are each independently selected fromhydrogen, —C₁-C₈ alkyl, —C₂-C₈ alkenyl, —C₂-C₈ alkynyl, substituted—C₁-C₈ alkyl, substituted —C₂-C₈ alkenyl, substituted —C₂-C₈ alkynyl,aryl, substituted aryl, heteroaryl, substituted heteroaryl,heterocyclic, substituted heterocyclic, —C₃-C₁₂ cycloalkyl, —C₃-C₁₂cycloalkenyl, substituted —C₃-C₁₂ cycloalkyl, or substituted —C₃-C₁₂cycloalkenyl.

In still another example, where Y₁-Y₃ are independently selected from—CR₇, N, NR₇, S and O, where R₇ is as previously defined above. A is—C(O)—O—R₁ or —C(O)—NH—R₁, where R₁ is —C₁-C₈ alkyl, —C₂-C₈ alkenyl,—C₂-C₈ alkynyl, substituted —C₁-C₈ alkyl, substituted —C₂-C₈ alkenyl,substituted —C₂-C₈ alkynyl, —C₃-C₁₂ cycloalkyl, —C₃-C₁₂ cycloalkenyl,substituted —C₃-C₁₂ cycloalkyl, or substituted —C₃-C₁₂ cycloalkenyl. Gis —NHSO₂—R₃, where R₃ is selected from aryl, substituted aryl,heteroaryl, substituted heteroaryl, heterocyclic, substitutedheterocyclic, —C₃-C₁₂ cycloalkyl, —C₃-C₁₂ cycloalkenyl, substituted—C₃-C₁₂ cycloalkyl, or substituted —C₃-C₁₂ cycloalkenyl.

In still yet another example, where Y₁-Y₃ are independently selectedfrom —CR₇, N, NR₇, S and O, where R₇ is as previously defined above. Ais —C(O)—O—R₁, where R₁ is —C₃-C₁₂ cycloalkyl or substituted —C₃-C₁₂cycloalkyl. G is —NHSO₂—R₃, where R₃ is selected from —C₃-C₁₂ cycloalkylor substituted —C₃-C₁₂ cycloalkyl.

In another example, where Y₁-Y₃ are independently selected from —CR₇, N,NR₇, S and O, where R₇ is as previously defined above. A is —C(O)—NH—R₁,where R₁ is —C₁-C₈ alkyl or substituted —C₁-C₈ alkyl. G is —NHSO₂—R₃,where R₃ is selected from —C₃-C₁₂ cycloalkyl or substituted —C₃-C₁₂cycloalkyl.

In one embodiment of the invention is a compound represented by FormulaVII:

or a pharmaceutically acceptable salt, ester or prodrug thereof, aloneor in combination with a pharmaceutically acceptable carrier orexcipient, where Y₁-Y₃ are independently selected selected from CR₇, N,NR₇, S and O, wherein R₇ is independently selected at each occurrencefrom:

-   -   (i) hydrogen; halogen; —NO₂; —CN;    -   (ii) —M—R₄, M is O, S, NH;    -   (iii) NR₄R₅;    -   (iv) —C₁-C₈ alkyl, —C₂-C₈ alkenyl, or —C₂-C₈ alkynyl each        containing 0, 1, 2, or 3 heteroatoms selected from O, S, or N;        substituted —C₁-C₈ alkyl, substituted —C₂-C₈ alkenyl, or        substituted —C₂-C₈ alkynyl each containing 0, 1, 2, or 3        heteroatoms selected from O, S or N; —C₃-C₁₂ cycloalkyl, or        substituted —C₃-C₁₂ cycloalkyl; —C₃-C₁₂ cycloalkenyl, or        substituted —C₃-C₁₂ cycloalkenyl;    -   (v) aryl; substituted aryl; heteroaryl; substituted heteroaryl;        and    -   (vi) heterocycloalkyl or substituted heterocycloalkyl;    -   A, G, R₄ and R₅ are are as defined in the first embodiment.

In one example, where Y₁-Y₃ are independently selected from —CR₇, N,NR₇, S and O, where R₇ is defined immediately above. A is selected fromthe group consisting of —C(O)—R₁, —C(O)—O—R₁ and —C(O)—NH—R₁, where R₁is selected from aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic, substituted heterocyclic, —C₁-C₈ alkyl, —C₂-C₈alkenyl, —C₂-C₈ alkynyl, substituted —C₁-C₈ alkyl, substituted —C₂-C₈alkenyl, substituted —C₂-C₈ alkynyl, —C₃-C₁₂ cycloalkyl, —C₃-C₁₂cycloalkenyl, substituted —C₃-C₁₂ cycloalkyl, or substituted —C₃-C₁₂cycloalkenyl. G can be —NH—SO₂—NR₄R₅ or —NHSO₂—R₃, where R₃ is selectedfrom —C₁-C₈ alkyl, —C₂-C₈ —C₂-C₈ alkynyl, aryl, substituted aryl,heteroaryl, substituted heteroaryl, heterocyclic, substitutedheterocyclic, —C₃-C₁₂ cycloalkyl, —C₃-C₁₂ cycloalkenyl, substituted—C₃-C₁₂ cycloalkyl, or substituted —C₃-C₁₂ cycloalkenyl, and R₄ and R₅are each independently selected from hydrogen, —C₁-C₈ alkyl, —C₂-C₈alkenyl, —C₂-C₈ alkynyl, substituted —C₁-C₈ alkyl, substituted —C₂-C₈alkenyl, substituted —C₂-C₈ alkynyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, heterocyclic, substituted heterocyclic, —C₃-C₁₂cycloalkyl, —C₃-C₁₂ cycloalkenyl, substituted —C₃-C₁₂ cycloalkyl, orsubstituted —C₃-C₁₂ cycloalkenyl.

In still another example, where Y₁-Y₃ are independently selected from—CR₇, N, NR₇, S and O, where R₇ is as previously defined above. A is—C(O)—O—R₁ or —C(O)—NH—R₁, where R₁ is —C₁-C₈ alkyl, —C₂-C₈ alkenyl,—C₂-C₈ alkynyl, substituted —C₁alkyl, substituted —C₂-C₈ alkenyl,substituted —C₂-C₈ alkynyl, —C₃-C₁₂ cycloalkyl, —C₃-C₁₂ cycloalkenyl,substituted —C₃-C₁₂ cycloalkyl, or substituted —C₃-C₁₂ cycloalkenyl. Gis —NHSO₂—R₃, where R₃ is selected from aryl, substituted aryl,heteroaryl, substituted heteroaryl, heterocyclic, substitutedheterocyclic, —C₃-C₁₂ cycloalkyl, —C₃-C₁₂ cycloalkenyl, substituted—C₃-C₁₂ cycloalkyl, or substituted —C₃-C₁₂ cycloalkenyl.

In still yet another example, where Y₁-Y₃ are independently selectedfrom —CR₇, N, NR₇, S and O, where R₇ is as previously defined above. Ais —C(O)—O—R₁, where R₁ is —C₃-C₁₂ cycloalkyl or substituted —C₃-C₁₂cycloalkyl. G is —NHSO₂—R₃, where R₃ is selected from —C₃-C₁₂ cycloalkylor substituted —C₃-C₁₂ cycloalkyl.

In another example, where Y₁-Y₃ are independently selected from —CR₇, N,NR₇, S and O, where R₇ is as previously defined above. A is —C(O)—NH—R₁,where R₁ is —C₁-C₈ alkyl or substituted —C₁-C₈ alkyl. G is —NHSO₂—R₃,where R₃ is selected from —C₃-C₁₂ cycloalkyl or substituted —C₃-C₁₂cycloalkyl.

Representative compounds of the invention include, but are not limitedto, the following compounds (Table 1) according to Formula IX:

TABLE 1 (IX)

Example # A Q G 22

23

24

25

26

27

28

29

30

31

32

33

34

35

36

37

38

39

40

41

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49

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51

52

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66

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68

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71

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100

101

102

103

104

105

106

107

108

109

110

The present invention also features pharmaceutical compositionscomprising a compound of the present invention, or a pharmaceuticallyacceptable salt, ester or prodrug thereof.

According to an alternate embodiment, the pharmaceutical compositions ofthe present invention may further contain other anti-HCV agents.Examples of anti-HCV agents include, but are not limited to, interferon(e.g., alpha-interferon, beta-interferon, consensus interferon,pegylated interferon, or albumin or other conjugated interferon),ribavirin, and amantadine. For further details see S. Tan, A. Pause, Y.Shi, N. Sonenberg, Hepatitis C Therapeutics: Current Status and EmergingStrategies, Nature Rev. Drug Discov., 1, 867-881 (2002); WO 00/59929(2000); WO 99/07733 (1999); WO 00/09543 (2000); WO 99/50230 (1999); U.S.Pat. No. 5,861,297 (1999); and US2002/0037998 (2002) which are hereinincorporated by reference in their entirety.

According to an additional embodiment, the pharmaceutical compositionsof the 1 5 present invention may further contain other HCV proteaseinhibitors.

According to yet another embodiment, the pharmaceutical compositions ofthe present invention may further comprise inhibitor(s) of other targetsin the HCV life cycle, including, but not limited to, helicase,polymerase, metalloprotease, and internal ribosome entry site (IRES).

According to another embodiment, the pharmaceutical compositions of thepresent invention may further comprise another anti-viral,anti-bacterial, anti-fungal or anti-cancer agent, or an immunemodulator, or another thearapeutic agent.

According to still another embodiment, the present invention includesmethods of treating hepatitis C infections in a subject in need of suchtreatment by administering to said subject an anti-HCV virally effectiveamount of a compound of the present invention or a pharmaceuticallyacceptable salt, ester, or prodrug thereof.

According to a further embodiment, the present invention includesmethods of treating hepatitis C infections in a subject in need of suchtreatment by administering to said subject an anti-HCV virally effectiveamount or an inhibitory amount of a pharmaceutical composition of thepresent invention.

An additional embodiment of the present invention includes methods oftreating biological samples by contacting the biological samples withthe compounds of the present invention.

Yet a further aspect of the present invention is a process of making anyof the compounds delineated herein employing any of the synthetic meansdelineated herein.

Definitions

Listed below are definitions of various terms used to describe thisinvention. These definitions apply to the terms as they are usedthroughout this specification and claims, unless otherwise limited inspecific instances, either individually or as part of a larger group.

The term “C₁-C₆ alkyl,” or “C₁-C₈ alkyl,” as used herein, refer tosaturated, straight- or branched-chain hydrocarbon radicals containingbetween one and six, or one and eight carbon atoms, respectively.Examples of C₁-C₆ alkyl radicals include, but are not limited to,methyl, ethyl, propyl, isopropyl, n-butyl, tert-butyl, neopentyl,n-hexyl radicals; and examples of C₁-C₈ alkyl radicals include, but arenot limited to, methyl, ethyl, propyl, isopropyl, n-butyl, tert-butyl,neopentyl, n-hexyl, heptyl, octyl radicals.

The term “C₂-C₆ alkenyl,” or “C₂-C₈ alkenyl,” as used herein, denote amonovalent group derived from a hydrocarbon moiety by the removal of asingle hydrogen atom wherein the hydrocarbon moiety has at least onecarbon-carbon double bond and contains from two to six, or two to eightcarbon atoms, respectively. Alkenyl groups include, but are not limitedto, for example, ethenyl, propenyl, butenyl, 1-methyl-2-buten-1-yl,heptenyl, octenyl and the like.

The term “C₂-C₆ alkynyl,” or “C₂-C₈ alkynyl,” as used herein, denote amonovalent group derived from a hydrocarbon moiety by the removal of asingle hydrogen atom wherein the hydrocarbon moiety has at least onecarbon-carbon triple bond and contains from two to six, or two to eightcarbon atoms, respectively. Representative alkynyl groups include, butare not limited to, for example, ethynyl, 1-propynyl, 1-butynyl,heptynyl, octynyl and the like.

The term “C₃-C₈-cycloalkyl”, or “C₃-C₁₂-cycloalkyl,” as used herein,denotes a monovalent group derived from a monocyclic or polycyclicsaturated carbocyclic ring compound by the removal of a single hydrogenatom where the saturated carbocyclic ring compound has from 3 ot 8, orfrom 3 to 12, ring atoms, respectively. Examples of C₃-C₈-cycloalkylinclude, but not limited to, cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, cyclopentyl and cyclooctyl; and examples ofC₃-C₁₂-cycloalkyl include, but not limited to, cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, bicyclo [2.2. 1] heptyl, and bicyclo [2.2.2]octyl.

The term “C₃-C₈-cycloalkenyl”, or “C₃-C₁₂-cycloalkenyl” as used herein,denote a monovalent group derived from a monocyclic or polycycliccarbocyclic ring compound having at least one carbon-carbon double bondby the removal of a single hydrogen atom where the carbocyclic ringcompound has from 3 ot 8, or from 3 to 12, ring atoms, respectively.Examples of C₃-C₈-cycloalkenyl include, but not limited to,cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl,cyclooctenyl, and the like; and examples of C₃-C₁₂-cycloalkenyl include,but not limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl,cyclohexenyl, cycloheptenyl, cyclooctenyl, and the like.

The term “aryl,” as used herein, refers to a mono- or bicycliccarbocyclic ring system having one or two aromatic rings including, butnot limited to, phenyl, naphthyl, tetrahydronaphthyl, indanyl, idenyland the like.

The term “arylalkyl,” as used herein, refers to a C₁-C₃ alkyl or C₁-C₆alkyl residue attached to an aryl ring. Examples include, but are notlimited to, benzyl, phenethyl and the like.

The term “heteroaryl,” as used herein, refers to a mono-, bi-, ortri-cyclic aromatic radical or ring having from five to ten ring atomsof which at least one ring atom is selected from S, O and N; wherein anyN or S contained within the ring may be optionally oxidized. Heteroarylincludes, but is not limited to, pyridinyl, pyrazinyl, pyrimidinyl,pyrrolyl, pyrazolyl, imidazolyl, thiazolyl, oxazolyl, isooxazolyl,thiadiazolyl, oxadiazolyl, thiophenyl, furanyl, quinolinyl,isoquinolinyl, benzimidazolyl, benzooxazolyl, quinoxalinyl, and thelike.

The term “heteroarylalkyl,” as used herein, refers to a C₁-C₃ alkyl orC₁-C₆ alkyl residue residue attached to a heteroaryl ring. Examplesinclude, but are not limited to, pyridinylmethyl, pyrimidinylethyl andthe like.

The terms “heterocyclic” and “heterocycloalkyl,” can be usedinterchangeably, and refer to a non-aromatic 3-, 4-, 5-, 6- or7-membered ring or a bi- or tri-cyclic group fused system, where (i)each ring contains between one and three heteroatoms independentlyselected from oxygen, sulfur and nitrogen, (ii) each 5-membered ring has0 to 1 double bonds and each 6-membered ring has 0 to 2 double bonds,(iii) the nitrogen and sulfur heteroatoms may optionally be oxidized,(iv) the nitrogen heteroatom may optionally be quaternized, and (iv) anyof the above rings may be fused to a benzene ring. Representativeheterocycloalkyl groups include, but are not limited to, [1,3]dioxolane,pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl,piperidinyl, piperazinyl, oxazolidinyl, isoxazolidinyl, morpholinyl,thiazolidinyl, isothiazolidinyl, and tetrahydrofuryl. Suchheterocycloalkyl groups may be further substituted to give substitutedheterocycloalkyl.

The term “substituted” as used herein, refers to independent replacementof one, two, or three or more of the hydrogen atoms thereon withsubstituents including, but not limited to, —F, —Cl, —Br, —I, —OH,protected hydroxy, —NO₂, —CN, —NH₂, protected amino, —NH —C₁C₁₂-alkyl,—NH —C₂-C₁₂-alkenyl, —NH —C₂-C₁₂-alkenyl, —NH —C₃-C₁₂-cycloalkyl,—NH-aryl, —NH-heteroaryl, —NH-heterocycloalkyl, -dialkylamino,-diarylamino, -diheteroarylamino, —O—C₁-C₁₂-alkyl, —O—C₂-C₁₂-alkenyl,—O—C₂-C₁₂-alkenyl, —O—C₃-C₁₂-cycloalkyl, —O-aryl, —O-heteroaryl,—O-heterocycloalkyl, —C(O)—C₁-C₁₂-alkyl, —C(O)—C₂-C₁₂-alkenyl,—C(O)—C₂-C₁₂-alkenyl, —C(O)—C₃-C₁₂-cycloalkyl, —C(O)-aryl,—C(O)-heteroaryl, —C(O)-heterocycloalkyl, —CONH₂, —CONH—C₁-C₁₂-alkyl,—CONH—C₂-C₁₂-alkenyl, —CONH—C₂-C₁₂-alkenyl, —CONH—C₃-C₁₂-cycloalkyl,—CONH-aryl, —CONH-heteroaryl, —CONH-heterocycloalkyl,—OCO₂—C₁-C₁₂-alkyl, —OCO₂—C₂-C₁₂-alkenyl, —OCO₂—C₂-C₁₂-alkenyl,—OCO₂-C₃-C₁₂-cycloalkyl, —OCO₂-aryl, —OCO₂-heteroaryl—OCO₂-heterocycloalkyl, —OCONH₂, —OCONH—C₁-C₁₂-alkyl,—OCONH—C₂-C₁₂-alkenyl, —OCONH—C₂-C₁₂-alkenyl, —OCONH—C₃-C₁₂-cycloalkyl,—OCONH-aryl, —OCONH-heteroaryl, —OCONH-heterocycloalkyl,—NHC(O)—C₁-C₁₂-alkyl, —NHC(O)—C₂-C₁₂-alkenyl, —NHC(O)—C₂-C ₁₂-alkenyl,—NHC(O)—C₃-C₁₂-cycloalkyl, —NHC(O)-aryl, —NHC(O)-heteroaryl,—NHC(O)-heterocycloalkyl, —NHCO₂—C₁-C₁₂-alkyl, —NHCO₂—C₂-C₁₂-alkenyl,—NHCO₂—C₂-C₁₂-alkenyl, —NHCO₂—C₃-C₁₂-cycloalkyl,—NHCO₂-aryl,—NHCO₂-heteroaryl, —NHCO₂-heterocycloalkyl, —NHC(O)NH₂,—NHC(O)NH—C₁-C₁₂-alkyl, —NHC(O)NH—C₂-C₁₂-alkenyl,—NHC(O)NH—C₂-C₁₂-alkenyl, —NHC(O)NH—C₃-C₁₂-cycloalkyl, —NHC(O)NH-aryl,—NHC(O)NH-heteroaryl, —NHC(O)NH-heterocycloalkyl, NHC(S)NH₂,—NHC(S)NH—C₁-C₁₂-alkyl, —NHC(S)NH—C₂-C₁₂-alkenyl,—NHC(S)NH—C₂-C₁₂-alkenyl, —NHC(S)NH—C₃-C₁₂-cycloalkyl, —NHC(S)NH-aryl,—NHC(S)NH-heteroaryl, —NHC(S)NH-heterocycloalkyl,—NHC(NH)NH₂—NHC(NH)NH—C₁-C₁₂-alkyl, —NHC(NH)NH—C₂-C₁₂-alkenyl,—NHC(NH)NH—C₂-C₁₂-alkenyl, —NHC(NH)NH—C₃-C₁₂-cycloalkyl,—NHC(NH)NH-aryl, —NHC(NH)NH-heteroaryl, —NHC(NH)NH-heterocycloalkyl,—NHC(NH)—C₁-C₁₂-alkyl, —NHC(NH)—C₂-C₁₂-alkenyl, —NHC(NH)—C₂-C₁₂-alkenyl,—NHC(NH)—C₃-C₁₂-cycloalkyl, —NHC(NH)-aryl, —NHC(NH)-heteroaryl,—NHC(NH)-heterocycloalkyl, —C(NH)NH—C₁-C₁₂-alkyl,—C(NH)NH—C₂-C₁₂-alkenyl, —C(NH)NH—C₂-C₁₂-alkenyl,—C(NH)NH—C₃-C₁₂-cycloalkyl, —C(NH)NH-aryl, —C(NH)NH-heteroaryl,—C(NH)NH-heterocycloalkyl, —S(O)—C₁-C₁₂-alkyl, —S(O)—C₂-C₁₂-alkenyl,—S(O)—C₂-C₁₂-alkenyl, —S(O)—C₃-C₁₂-cycloalkyl, —S(O)-aryl,—S(O)-heteroaryl, —S(O)-heterocycloalkyl —SO₂NH₂, —SO₂NH—C₁-C₁₂-alkyl,-SO₂NH—C₂-C₁₂-alkenyl, —SO₂NH—C₂-C₁₂-alkenyl, —SO₂NH—C₃-C₁₂-cycloalkyl,—SO₂NH-aryl, —SO₂NH-heteroaryl, —SO₂NH-heterocycloalkyl,—NHSO₂-C₁-C₁₂-alkyl, —NHSO₂-C₂-C₁₂-alkenyl, —NHSO₂—C₂-C₁₂-alkenyl,—NHSO₂—C₃-C₁₂-cycloalkyl, —NHSO₂-aryl, —NHSO₂-heteroaryl,—NHSO₂-heterocycloalkyl, —CH₂NH₂, —CH₂SO₂CH₃, -aryl, -arylalkyl,-heteroaryl, -heteroarylalkyl, -heterocycloalkyl, —C₃-C₁₂-cycloalkyl,polyalkoxyalkyl, polyalkoxy, -methoxymethoxy, -methoxyethoxy, —SH,—S—C₁-C₁₂-alkyl, —S—C₂-C₁₂-alkenyl, —S—C₂-C₁₂-alkenyl,—S—C₃-C₁₂-cycloalkyl, —S-aryl, —S-heteroaryl, —S-heterocycloalkyl, ormethylthiomethyl. It is understood that the aryls, heteroaryls, alkyls,and the like can be further substituted. In some cases, each substituentin a substituted moiety is additionally optionally substituted with oneor more groups, each group being independently selected from —F, —Cl,—Br, —I, —OH, —NO₂, —CN, or —NH₂.

In accordance with the invention, any of the aryls, substituted aryls,heteroaryls and substituted heteroaryls described herein, can be anyaromatic group. Aromatic groups can be substituted or unsubstituted.

It is understood that any alkyl, alkenyl, alkynyl, cycloalkyl andcycloalkenyl moiety described herein can also be an aliphatic group, analicyclic group or a heterocyclic group. An “aliphatic group” isnon-aromatic moiety that may contain any combination of carbon atoms,hydrogen atoms, halogen atoms, oxygen, nitrogen or other atoms, andoptionally contain one or more units of unsaturation, e.g., doubleand/or triple bonds. An aliphatic group may be straight chained,branched or cyclic and preferably contains between about 1 and about 24carbon atoms, more typically between about 1 and about 12 carbon atoms.In addition to aliphatic hydrocarbon groups, aliphatic groups include,for example, polyalkoxyalkyls, such as polyalkylene glycols, polyamines,and polyimines, for example. Such aliphatic groups may be furthersubstituted. It is understood that aliphatic groups may be used in placeof the alkyl, alkenyl, alkynyl, alkylene, alkenylene, and alkynylenegroups described herein.

The term “alicyclic,” as used herein, denotes a monovalent group derivedfrom a monocyclic or polycyclic saturated carbocyclic ring compound bythe removal of a single hydrogen atom. Examples include, but not limitedto, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, bicyclo [2.2.1]heptyl, and bicyclo [2.2.2] octyl. Such alicyclic groups may be furthersubstituted.

It will be apparent that in various embodiments of the invention, thesubstituted or unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl,cycloalkenyl, cycloalkynyl, arylalkyl, heteroarylalkyl, andheterocycloalkyl are intended to be monovalent or divalent. Thus,alkylene, alkenylene, and alkynylene, cycloaklylene, cycloalkenylene,cycloalkynylene, arylalkylene, hetoerarylalkylene andheterocycloalkylene groups are to be included in the above definitions,and are applicable to provide the formulas herein with proper valency.

The term “hydroxy activating group”, as used herein, refers to a labilechemical moiety which is known in the art to activate a hydroxy group sothat it will depart during synthetic procedures such as in asubstitution or elimination reactions. Examples of hydroxy activatinggroup include, but not limited to, mesylate, tosylate, triflate,p-nitrobenzoate, phosphonate and the like.

The term “activated hydroxy”, as used herein, refers to a hydroxy groupactivated with a hydroxy activating group, as defined above, includingmesylate, tosylate, triflate, p-nitrobenzoate, phosphonate groups, forexample.

The term “protected hydroxy,” as used herein, refers to a hydroxy groupprotected with a hydroxy protecting group, as defined above, includingbenzoyl, acetyl, trimethylsilyl, triethylsilyl, methoxymethyl groups,for example.

The terms “halo” and “halogen,” as used herein, refer to an atomselected from fluorine, chlorine, bromine and iodine.

The compounds described herein contain one or more asymmetric centersand thus give rise to enantiomers, diastereomers, and otherstereoisomeric forms that may be defined, in terms of absolutestereochemistry, as (R)— or (S)—, or as (D)- or (L)- for amino acids.The present invention is meant to include all such possible isomers, aswell as their racemic and optically pure forms. Optical isomers may beprepared from their respective optically active precursors by theprocedures described above, or by resolving the racemic mixtures. Theresolution can be carried out in the presence of a resolving agent, bychromatography or by repeated crystallization or by some combination ofthese techniques which are known to those skilled in the art. Furtherdetails regarding resolutions can be found in Jacques, et al.,Enantiomers, Racemates, and Resolutions (John Wiley & Sons, 1981). Whenthe compounds described herein contain olefinic double bonds or othercenters of geometric asymmetry, and unless specified otherwise, it isintended that the compounds include both E and Z geometric isomers.Likewise, all tautomeric forms are also intended to be included. Theconfiguration of any carbon-carbon double bond appearing herein isselected for convenience only and is not intended to designate aparticular configuration unless the text so states; thus a carbon-carbondouble bond depicted arbitrarily herein as trans may be cis, trans, or amixture of the two in any proportion.

The term “subject” as used herein refers to a mammal. A subjecttherefore refers to, for example, dogs, cats, horses, cows, pigs, guineapigs, and the like. Preferably the subject is a human. When the subjectis a human, the subject may be referred to herein as a patient.

As used herein, the term “pharmaceutically acceptable salt” refers tothose salts of the compounds formed by the process of the presentinvention which are, within the scope of sound medical judgment,suitable for use in contact with the tissues of humans and lower animalswithout undue toxicity, irritation, allergic response and the like, andare commensurate with a reasonable benefit/risk ratio. Pharmaceuticallyacceptable salts are well known in the art.

Berge, et al. describes pharmaceutically acceptable salts in detail inJ. Pharmaceutical Sciences, 66:1-19 (1977). The salts can be prepared insitu during the final isolation and purification of the compounds of theinvention, or separately by reacting the free base function with asuitable organic acid. Examples of pharmaceutically acceptable include,but are not limited to, nontoxic acid addition salts are salts of anamino group formed with inorganic acids such as hydrochloric acid,hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid orwith organic acids such as acetic acid, maleic acid, tartaric acid,citric acid, succinic acid or malonic acid or by using other methodsused in the art such as ion exchange. Other pharmaceutically acceptablesalts include, but are not limited to, adipate, alginate, ascorbate,aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate,camphorate, camphorsulfonate, citrate, cyclopentanepropionate,digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate,glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate,hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate,lactate, laurate, lauryl sulfate, malate, maleate, malonate,methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate,oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate,phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate,tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts,and the like. Representative alkali or alkaline earth metal saltsinclude sodium, lithium, potassium, calcium, magnesium, and the like.Further pharmaceutically acceptable salts include, when appropriate,nontoxic ammonium, quaternary ammonium, and amine cations formed usingcounterions such as halide, hydroxide, carboxylate, sulfate, phosphate,nitrate, alkyl having from 1 to 6 carbon atoms, sulfonate and arylsulfonate.

The term “hydroxy protecting group,” as used herein, refers to a labilechemical moiety which is known in the art to protect a hydroxy groupagainst undesired reactions during synthetic procedures. After saidsynthetic procedure(s) the hydroxy protecting group as described hereinmay be selectively removed. Hydroxy protecting groups as known in theare described generally in T. H. Greene and P. G., S.Wuts, ProtectiveGroups in Organic Synthesis, 3rd edition, John Wiley & Sons, New York(1999). Examples of hydroxy protecting groups include benzyloxycarbonyl,4-nitrobenzyloxycarbonyl, 4-bromobenzyloxycarbonyl,4-methoxybenzyloxycarbonyl, methoxycarbonyl, tert-butoxycarbonyl,isopropoxycarbonyl, diphenylmethoxycarbonyl,2,2,2-trichloroethoxycarbonyl, 2-(trimethylsilyl)ethoxycarbonyl,2-furfuryloxycarbonyl, allyloxycarbonyl, acetyl, formyl, chloroacetyl,trifluoroacetyl, methoxyacetyl, phenoxyacetyl, benzoyl, methyl, t-butyl,2,2,2-trichloroethyl, 2-trimethylsilyl ethyl, 1,1-dimethyl-2-propenyl,3-methyl-3-butenyl, allyl, benzyl, para-methoxybenzyldiphenylmethyl,triphenylmethyl (trityl), tetrahydrofuryl, methoxymethyl,methylthiomethyl, benzyloxymethyl, 2,2,2-triehloroethoxymethyl,2-(trimethylsilyl)ethoxymethyl, methanesulfonyl, para-toluenesulfonyl,trimethylsilyl, triethylsilyl, triisopropylsilyl, and the like.Preferred hydroxy protecting groups for the present invention are acetyl(Ac or —C(O)CH₃), benzoyl (Bz or —C(O)C₆H₅), and trimethylsilyl (TMS or—Si(CH₃)₃).

The term “amino protecting group,” as used herein, refers to a labilechemical moiety which is known in the art to protect an amino groupagainst undesired reactions during synthetic procedures. After saidsynthetic procedure(s) the amino protecting group as described hereinmay be selectively removed. Amino protecting groups as known in the aredescribed generally in T. H. Greene and P. G. M. Wuts, Protective Groupsin Organic Synthesis, 3rd edition, John Wiley & Sons, New York (1999).Examples of amino protecting groups include, but are not limited to,t-butoxycarbonyl, 9-fluorenylmethoxycarbonyl, benzyloxycarbonyl, and thelike.

As used herein, the term “pharmaceutically acceptable ester” refers toesters of the compounds formed by the process of the present inventionwhich hydrolyze in vivo and include those that break down readily in thehuman body to leave the parent compound or a salt thereof. Suitableester groups include, for example, those derived from pharmaceuticallyacceptable aliphatic carboxylic acids, particularly alkanoic, alkenoic,cycloalkanoic and alkanedioic acids, in which each alkyl or alkenylmoiety advantageously has not more than 6 carbon atoms. Examples ofparticular esters include, but are not limited to, formates, acetates,propionates, butyrates, acrylates and ethylsuccinates.

The term “pharmaceutically acceptable prodrugs” as used herein refers tothose prodrugs of the compounds formed by the process of the presentinvention which are, within the scope of sound medical judgment,suitable for use in contact with the tissues of humans and lower animalswith undue toxicity, irritation, allergic response, and the like,commensurate with a reasonable benefit/risk ratio, and effective fortheir intended use, as well as the zwitterionic forms, where possible,of the compounds of the present invention. “Prodrug”, as used hereinmeans a compound which is convertible in vivo by metabolic means (e.g.by hydrolysis) to afford any compound delineated by the formulae of theinstant invention. Various forms of prodrugs are known in the art, forexample, as discussed in Bundgaard, (ed.), Design of Prodrugs, Elsevier(1985); Widder, et al. (ed.), Methods in Enzymology, vol. 4, AcademicPress (1985); Krogsgaard-Larsen, et al., (ed). “Design and Applicationof Prodrugs, Textbook of Drug Design and Development, Chapter 5, 113-191(1991); Bundgaard, et al., Journal of Drug Deliver Reviews,8:1-38(1992); Bundgaard, J. of Pharmaceutical Sciences, 77:285 et seq.(1988); Higuchi and Stella (eds.) Prodrugs as Novel Drug DeliverySystems, American Chemical Society (1975); and Bernard Testa & JoachimMayer, “Hydrolysis In Drug And Prodrug Metabolism: Chemistry,Biochemistry And Enzymology,” John Wiley and Sons, Ltd. (2002).

The term “acyl” includes residues derived from acids, including but notlimited to carboxylic acids, carbamic acids, carbonic acids, sulfonicacids, and phosphorous acids. Examples include aliphatic carbonyls,aromatic carbonyls, aliphatic sulfonyls, aromatic sulfinyls, aliphaticsulfinyls, aromatic phosphates and aliphatic phosphates. Examples ofaliphatic carbonyls include, but are not limited to, acetyl, propionyl,2-fluoroacetyl, butyryl, 2-hydroxy acetyl, and the like.

The term “aprotic solvent,” as used herein, refers to a solvent that isrelatively inert to proton activity, i.e., not acting as a proton-donor.Examples include, but are not limited to, hydrocarbons, such as hexaneand toluene, for example, halogenated hydrocarbons, such as, forexample, methylene chloride, ethylene chloride, chloroform, and thelike, heterocyclic compounds, such as, for example, tetrahydrofuran andN-methylpyrrolidinone, and ethers such as diethyl ether,bis-methoxymethyl ether. Such solvents are well known to those skilledin the art, and individual solvents or mixtures thereof may be preferredfor specific compounds and reaction conditions, depending upon suchfactors as the solubility of reagents, reactivity of reagents andpreferred temperature ranges, for example. Further discussions ofaprotic solvents may be found in organic chemistry textbooks or inspecialized monographs, for example: Organic Solvents PhysicalProperties and Methods of Purification, 4th ed., edited by John A.Riddick et al., Vol. II, in the Techniques of Chemistry Series, JohnWiley & Sons, NY, 1986.

The terms “protogenic organic solvent,” or “protic solvent” as usedherein, refer to a solvent that tends to provide protons, such as analcohol, for example, methanol, ethanol, propanol, isopropanol, butanol,t-butanol, and the like. Such solvents are well known to those skilledin the art, and individual solvents or mixtures thereof may be preferredfor specific compounds and reaction conditions, depending upon suchfactors as the solubility of reagents, reactivity of reagents andpreferred temperature ranges, for example. Further discussions ofprotogenic solvents may be found in organic chemistry textbooks or inspecialized monographs, for example: Organic Solvents PhysicalProperties and Methods of Purification, 4th ed., edited by John A.Riddick et al, Vol. II, in the Techniques of Chemistry Series, JohnWiley & Sons, NY, 1986.

Combinations of substituents and variables envisioned by this inventionare only those that result in the formation of stable compounds. Theterm “stable”, as used herein, refers to compounds which possessstability sufficient to allow manufacture and which maintains theintegrity of the compound for a sufficient period of time to be usefulfor the purposes detailed herein (e.g., therapeutic or prophylacticadministration to a subject).

The synthesized compounds can be separated from a reaction mixture andfurther purified by a method such as column chromatography, highpressure liquid chromatography, or recrystallization. As can beappreciated by the skilled artisan, further methods of synthesizing thecompounds of the formulae herein will be evident to those of ordinaryskill in the art. Additionally, the various synthetic steps may beperformed in an alternate sequence or order to give the desiredcompounds. In addition, the solvents, temperatures, reaction durations,etc. delineated herein are for purposes of illustration only and one ofordinary skill in the art will recognize that variation of the reactionconditions can produce the desired bridged macrocyclic products of thepresent invention. Synthetic chemistry transformations and protectinggroup methodologies (protection and deprotection) useful in synthesizingthe compounds described herein are known in the art and include, forexample, those such as described in R. Larock, Comprehensive OrganicTransformations, VCH Publishers (1989); T. W. Greene and P. G. M. Wuts,Protective Groups in Organic Synthesis, 2d. E d., John Wiley and Sons(1991); L. Fieser and M. Fieser, Fieser and Fieser's Reagents forOrganic Synthesis, John Wiley and Sons (1994); and L. Paquette, ed.,Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons(1995).

The compounds of this invention may be modified by appending variousfunctionalities via any synthetic means delineated herein to enhanceselective biological properties. Such modifications are known in the artand include those which increase biological penetration into a givenbiological system (e.g., blood, lymphatic system, central nervoussystem), increase oral availability, increase solubility to allowadministration by injection, alter metabolism and alter rate ofexcretion.

Pharmaceutical Compostion

The pharmaceutical compositions of the present invention comprise atherapeutically effective amount of a compound of the present inventionformulated together with one or more pharmaceutically acceptablecarriers. As used herein, the term “pharmaceutically acceptable carrier”means a non-toxic, inert solid, semi-solid or liquid filler, diluent,encapsulating material or formulation auxiliary of any type. Someexamples of materials which can serve as pharmaceutically acceptablecarriers are sugars such as lactose, glucose and sucrose; starches suchas corn starch and potato starch; cellulose and its derivatives such assodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate;powdered tragacanth; malt; gelatin; talc; excipients such as cocoabutter and suppository waxes; oils such as peanut oil, cottonseed oil;safflower oil; sesame oil; olive oil; corn oil and soybean oil; glycols;such a propylene glycol; esters such as ethyl oleate and ethyl laurate;agar; buffering agents such as magnesium hydroxide and aluminumhydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer'ssolution; ethyl alcohol, and phosphate buffer solutions, as well asother non-toxic compatible lubricants such as sodium lauryl sulfate andmagnesium stearate, as well as coloring agents, releasing agents,coating agents, sweetening, flavoring and perfuming agents,preservatives and antioxidants can also be present in the composition,according to the judgment of the formulator. The pharmaceuticalcompositions of this invention can be administered to humans and otheranimals orally, rectally, parenterally, intracisternally,intravaginally, intraperitoneally, topically (as by powders, ointments,or drops), buccally, or as an oral or nasal spray.

The pharmaceutical compositions of this invention may be administeredorally, parenterally, by inhalation spray, topically, rectally, nasally,buccally, vaginally or via an implanted reservoir, preferably by oraladministration or administration by injection. The pharmaceuticalcompositions of this invention may contain any conventional non-toxicpharmaceutically-acceptable carriers, adjuvants or vehicles. In somecases, the pH of the formulation may be adjusted with pharmaceuticallyacceptable acids, bases or buffers to enhance the stability of theformulated compound or its delivery form. The term parenteral as usedherein includes subcutaneous, intracutaneous, intravenous,intramuscular, intraarticular, intraarterial, intrasynovial,intrasternal, intrathecal, intralesional and intracranial injection orinfusion techniques.

Liquid dosage forms for oral administration include pharmaceuticallyacceptable emulsions, microemulsions, solutions, suspensions, syrups andelixirs. In addition to the active compounds, the liquid dosage formsmay contain inert diluents commonly used in the art such as, forexample, water or other solvents, solubilizing agents and emulsifierssuch as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethylacetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyleneglycol, dimethylformamide, oils (in particular, cottonseed, groundnut,corn, germ, olive, castor, and sesame oils), glycerol,tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid estersof sorbitan, and mixtures thereof. Besides inert diluents, the oralcompositions can also include adjuvants such as wetting agents,emulsifying and suspending agents, sweetening, flavoring, and perfumingagents.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions may be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation may also be a sterile injectablesolution, suspension or emulsion in a nontoxic parenterally acceptablediluent or solvent, for example, as a solution in 1,3-butanediol. Amongthe acceptable vehicles and solvents that may be employed are water,Ringer's solution, U.S.P. and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil can beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectables.

The injectable formulations can be sterilized, for example, byfiltration through a bacterial-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved or dispersed in sterile water or other sterile injectablemedium prior to use.

In order to prolong the effect of a drug, it is often desirable to slowthe absorption of the drug from subcutaneous or intramuscular injection.This may be accomplished by the use of a liquid suspension ofcrystalline or amorphous material with poor water solubility. The rateof absorption of the drug then depends upon its rate of dissolutionwhich, in turn, may depend upon crystal size and crystalline form.Alternatively, delayed absorption of a parenterally administered drugform is accomplished by dissolving or suspending the drug in an oilvehicle. Injectable depot forms are made by forming microencapsulematrices of the drug in biodegradable polymers such aspolylactide-polyglycolide. Depending upon the ratio of drug to polymerand the nature of the particular polymer employed, the rate of drugrelease can be controlled. Examples of other biodegradable polymersinclude poly(orthoesters) and poly(anhydrides). Depot injectableformulations are also prepared by entrapping the drug in liposomes ormicroemulsions which are compatible with body tissues.

Compositions for rectal or vaginal administration are preferablysuppositories which can be prepared by mixing the compounds of thisinvention with suitable non-irritating excipients or carriers such ascocoa butter, polyethylene glycol or a suppository wax which are solidat ambient temperature but liquid at body temperature and therefore meltin the rectum or vaginal cavity and release the active compound.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activecompound is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier such as sodium citrate or dicalcium phosphateand/or: a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol, and silicic acid, b) binders such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,sucrose, and acacia, c) humectants such as glycerol, d) disintegratingagents such as agar-agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate, e) solutionretarding agents such as paraffin, f) absorption accelerators such asquaternary ammonium compounds, g) wetting agents such as, for example,cetyl alcohol and glycerol monostearate, h) absorbents such as kaolinand bentonite clay, and i) lubricants such as talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof In the case of capsules, tablets and pills, thedosage form may also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using such excipients as lactoseor milk sugar as well as high molecular weight polyethylene glycols andthe like.

The active compounds can also be in micro-encapsulated form with one ormore excipients as noted above. The solid dosage forms of tablets,dragees, capsules, pills, and granules can be prepared with coatings andshells such as enteric coatings, release controlling coatings and othercoatings well known in the pharmaceutical formulating art. In such soliddosage forms the active compound may be admixed with at least one inertdiluent such as sucrose, lactose or starch. Such dosage forms may alsocomprise, as is normal practice, additional substances other than inertdiluents, e.g., tableting lubricants and other tableting aids such amagnesium stearate and microcrystalline cellulose. In the case ofcapsules, tablets and pills, the dosage forms may also comprisebuffering agents. They may optionally contain opacifying agents and canalso be of a composition that they release the active ingredient(s)only, or preferentially, in a certain part of the intestinal tract,optionally, in a delayed manner. Examples of embedding compositionswhich can be used include polymeric substances and waxes.

Dosage forms for topical or transdermal administration of a compound ofthis invention include ointments, pastes, creams, lotions, gels,powders, solutions, sprays, inhalants or patches. The active componentis admixed under sterile conditions with a pharmaceutically acceptablecarrier and any needed preservatives or buffers as may be required.Ophthalmic formulation, ear drops, eye ointments, powders and solutionsare also contemplated as being within the scope of this invention.

The ointments, pastes, creams and gels may contain, in addition to anactive compound of this invention, excipients such as animal andvegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulosederivatives, polyethylene glycols, silicones, bentonites, silicic acid,talc and zinc oxide, or mixtures thereof.

Powders and sprays can contain, in addition to the compounds of thisinvention, excipients such as lactose, talc, silicic acid, aluminumhydroxide, calcium silicates and polyamide powder, or mixtures of thesesubstances. Sprays can additionally contain customary propellants suchas chlorofluorohydrocarbons.

Transdermal patches have the added advantage of providing controlleddelivery of a compound to the body. Such dosage forms can be made bydissolving or dispensing the compound in the proper medium. Absorptionenhancers can also be used to increase the flux of the compound acrossthe skin. The rate can be controlled by either providing a ratecontrolling membrane or by dispersing the compound in a polymer matrixor gel.

Antiviral Activity

An inhibitory amount or dose of the compounds of the present inventionmay range from about 0. 1 mg/Kg to about 500 mg/Kg, alternatively fromabout 1 to about 50 mg/Kg. Inhibitory amounts or doses will also varydepending on route of administration, as well as the possibility ofco-usage with other agents.

According to the methods of treatment of the present invention, viralinfections are treated or prevented in a subject such as a human orlower mammal by administering to the subject an anti-hepatitis C virallyeffective amount or an inhibitory amount of a compound of the presentinvention, in such amounts and for such time as is necessary to achievethe desired result. An additional method of the present invention is thetreatment of biological samples with an inhibitory amount of a compoundof composition of the present invention in such amounts and for suchtime as is necessary to achieve the desired result.

The term “anti-hepatitis C virally effective amount” of a compound ofthe invention, as used herein, mean a sufficient amount of the compoundso as to decrease the viral load in a biological sample or in a subject.As well understood in the medical arts, an anti-hepatitis C virallyeffective amount of a compound of this invention will be at a reasonablebenefit/risk ratio applicable to any medical treatment.

Lower or higher doses than those recited above may be required. Specificdosage and treatment regimens for any particular patient will dependupon a variety of factors, including the activity of the specificcompound employed, the age, body weight, general health status, sex,diet, time of administration, rate of excretion, drug combination, theseverity and course of the disease, condition or symptoms, the patient'sdisposition to the disease, condition or symptoms, and the judgment ofthe treating physician.

The term “inhibitory amount” of a compound of the present inventionmeans a sufficient amount to decrease the hepatitis C viral load in abiological sample or a subject. It is understood that when saidinhibitory amount of a compound of the present invention is administeredto a subject it will be at a reasonable benefit/risk ratio applicable toany medical treatment as determined by a physician. The term “biologicalsample(s),” as used herein, means a substance of biological originintended for administration to a subject. Examples of biological samplesinclude, but are not limited to, blood and components thereof such asplasma, platelets, subpopulations of blood cells and the like; organssuch as kidney, liver, heart, lung, and the like; sperm and ova; bonemarrow and components thereof, or stem cells. Thus, another embodimentof the present invention is a method of treating a biological sample bycontacting said biological sample with an inhibitory amount of acompound or pharmaceutical composition of the present invention.

Upon improvement of a subject's condition, a maintenance dose of acompound, composition or combination of this invention may beadministered, if necessary. Subsequently, the dosage or frequency ofadministration, or both, may be reduced, as a function of the symptoms,to a level at which the improved condition is retained when the symptomshave been alleviated to the desired level, treatment should cease. Thesubject may, however, require intermittent treatment on a long-termbasis upon any recurrence of disease symptoms.An additional method ofthe present invention is the treatment of biological samples with aninhibitory amount of a compound of the present invention in such amountsand for such time as is necessary to inhibit viral replication and/orreduce viral load. The term “inhibitory amount” means a sufficientamount to inhibit viral replication and/or decrease the hepatitis Cviral load in a biological sample. The term “biological sample(s)” asused herein means a substance of biological origin intended foradministration to a subject. Examples of biological samples include, butare not limited to blood and components thereof such as plasma,platelets, subpopulations of blood cells and the like; organs such askidney, liver, heart, lung, and the like; sperm and ova; bone marrow andcomponents thereof, or stem cells. Thus another embodiment of thepresent invention is a method of treating a biological sample bycontacting said biological sample with an inhibitory amount of acompound or pharmaceutical composition of the present invention.

It will be understood, however, that the total daily usage of thecompounds and compositions of the present invention will be decided bythe attending physician within the scope of sound medical judgment. Thespecific inhibitory dose for any particular patient will depend upon avariety of factors including the disorder being treated and the severityof the disorder; the activity of the specific compound employed; thespecific composition employed; the age, body weight, general health, sexand diet of the patient; the time of administration, route ofadministration, and rate of excretion of the specific compound employed;the duration of the treatment; drugs used in combination or coincidentalwith the specific compound employed; and like factors well known in themedical arts.

The total daily inhibitory dose of the compounds of this inventionadministered to a subject in single or in divided doses can be inamounts, for example, from 0.01 to 50 mg/kg body weight or more usuallyfrom 0. 1 to 25 mg/kg body weight. Single dose compositions may containsuch amounts or submultiples thereof to make up the daily dose. Ingeneral, treatment regimens according to the present invention compriseadministration to a patient in need of such treatment from about 10 mgto about 1000 mg of the compound(s) of this invention per day in singleor multiple doses.

Unless otherwise defined, all technical and scientific terms used hereinare accorded the meaning commonly known to one with ordinary skill inthe art. All publications, patents, published patent applications, andother references mentioned herein are hereby incorporated by referencein their entirety.

Abbreviations

Abbreviations which have been used in the descriptions of the schemesand the examples that follow are:

-   -   ACN for acetonitrile;    -   Ac for acetyl;    -   Boc for tert-butoxycarbonyl;    -   Bz for benzoyl;    -   Bn for benzyl;    -   CDI for carbonyldiimidazole;    -   dba for dibenzylidene acetone;    -   CDI for 1,1′-carbonyldiimidizole;    -   DBU for 1,8-diazabicyclo[5.4.0]undec-7-ene;    -   DCM for dichloromethane;    -   DIAD for diisopropylazodicarboxylate;    -   DMAP for dimethylaminopyridine;    -   DMF for dimethyl formamide;    -   DMSO for dimethyl sulfoxide;    -   dppb for diphenylphosphino butane;    -   EtOAc for ethyl acetate;    -   HATU for        2-(7-Aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium        hexafluorophosphate;    -   iPrOH for isopropanol;    -   NaHMDS for sodium bis(trimethylsilyl)amide;    -   NMO for N-methylmorpho line N-oxide;    -   MeOH for methanol;    -   Ph for phenyl;    -   POPd for dihydrogen        dichlorobis(di-tert-butylphosphino)palladium(II);    -   TBAHS for tetrabutyl ammonium hydrogen sulfate;    -   TEA for triethylamine;    -   THF for tetrahydrofuran;    -   TPP for triphenylphosphine;    -   Tris for Tris(hydroxymethyl)aminomethane;    -   BME for 2-mercaptoethanol;    -   BOP for benzotriazol-1-yloxy-tris(dimethylamino)phosphonium        hexafluorophosphate;    -   COD for cyclooctadiene;    -   DAST for diethylaminosulfur trifluoride;    -   DABCYL for        6-(N-4'-carboxy-4-(dimethylamino)azobenzene)-aminohexyl-1-O-(2-cyanoethyl)-(N,N-diisopropyl)-phosphoramidite;    -   DCM for dichloromethane;    -   DIAD for diisopropyl azodicarboxylate;    -   DIBAL-H for diisobutylaluminum hydride;    -   DIEA for diisopropyl ethylamine;    -   DMAP for N,N-dimethylaminopyridine;    -   DME for ethylene glycol dimethyl ether;    -   DMEM for Dulbecco's Modified Eagles Media;    -   DMF for N,N-dimethyl formamide;    -   DMSO for dimethylsulfoxide;    -   DUPHOS for

-   -   EDANS for 5-(2-Amino-ethylamino)-naphthalene-1-sulfonic acid;    -   EDCI or EDC for 1-(3-diethylaminopropyl)-3-ethylcarbodiimide        hydrochloride;    -   EtOAc for ethyl acetate;    -   HATU for O        (7-Azabenzotriazole-1-yl)-N,N,N′,N′-tetramethyluronium        hexafluorophosphate;    -   Hoveyda's Cat. for Dichloro(o-isopropoxyphenylmethylene)        (tricyclohexylphosphine)ruthenium(II);    -   KHMDS is potassium bis(trimethylsilyl) amide;    -   Ms for mesyl;    -   EtOAc for ethyl acetate;    -   g for gram(s);    -   h for hour(s);    -   NMM for N-4-methylmorpholine    -   PyBrOP for Bromo-tri-pyrolidino-phosphonium hexafluorophosphate;    -   Ph for phenyl;    -   RCM for ring-closing metathesis;    -   RT for room temperature    -   HATU for        O-(7-Azabenzotriazole-1-yl)-N,N,N′,N′-tetramethyluronium        hexafluoro-phosphate;    -   HPLC for high-performance liquid chromatography;    -   Ph for phenyl;    -   Me for methyl;    -   RT for reverse transcription;    -   RT-PCR for reverse transcription-polymerase chain reaction;    -   TEA for triethyl amine;    -   TFA for trifluoroacetic acid;    -   MeOH for methanol;    -   mg for milligram(s);    -   min for minute(s);    -   MS for mass spectrometry;    -   NMR for nuclear magnetic resonance;    -   rt for room temperature;    -   THF for tetrahydrofuran;    -   TLC for thin layer chromatography;    -   TPP or PPh₃ for triphenylphosphine;    -   tBOC or Boc for tert-butyloxy carbonyl; and    -   Xantphos for        4,5-Bis-diphenylphosphanyl-9,9-dimethyl-9H-xanthene.

Synthetic Methods

The compounds and processes of the present invention will be betterunderstood in connection with the following synthetic schemes thatillustrate the methods by which the compounds of the invention may beprepared.

Scheme 1 describes the synthesis of intermediate Ig. The cyclic peptideprecursor Ig was synthesized from Boc-L-2-amino-8-nonenoic acid Ia andcis-L-hydroxyproline methyl ester Ib via steps A-D set forth generallyin Scheme 1. For further details of the synthetic methods employed toproduce the cyclic peptide precursor Ig, see U.S. Pat. No. 6,608,027,which is herein incorporated by reference in its entirety. Other aminoacid derivatives containing a terminal alkene may be used in place of lain order to create varied macrocyclic structures (for further detailssee WO/0059929). Ring closure methathesis with a Ruthenium-basedcatalyst gave the desired key intermediate Ig (for further details onring closing metathesis see recent reviews: Grubbs et al., Acc. Chem.Res., 1995, 28, 446; Shrock et al., Tetrahedron 1999, 55, 8141;Furstner, A. Angew. Chem. Int. Ed. 2000, 39, 3012; Tmka et al., Acc.Chem. Res. 2001, 34, 18; and Hoveyda et al., Chem. Eur. J. 2001, 7,945).

Scheme 2 illustrates the general synthetic method of triazole analogs.Triazoles (2-2) were synthesized from alkynes (2-1) with TMSN₃, but notlimited to TMSN₃. The alkynes (2-1) are commercially available or madeby the Sonogashira reaction with primary alkyne (2-8) and aryl halide(2-9). For further details of the Sonogashira reaction see: Sonogashira,Comprehensive Organic Synthesis, Volume 3, Chapters 2,4 and Sonogashira,Synthesis 1977, 777. Intermediate (2-4) and (2-5) can be made throughSN2 replacement of activated hydroxyl group by converting hydroxyintermediate Ig to a suitable leaving group such as, but not limited toOMs, OTs, OTf, bromide, or iodide. Subsequent hydrolysis of the estergives compounds of formula (2-6) or (2-7).

Intermediate (3-1) was synthesized under the conditions with macrocyclicmesylate (2-3) and triazoles (2-2) as described in Scheme 2.Intermediate (3-1) may then undergo Suzuki coupling reactions,Sonogashira reactions, or Stille couplings at the position occupied bythe halide or OTf For further details concerning the Suzuki couplingreaction see: A. Suzuki, Pure Appl. Chem. 1991, 63, 419-422 and A. R.Martin, Y. Yang, Acta Chem. Scand. 1993, 47, 221-230. For furtherdetails of the Sonogashira reaction see: Sonogashira, ComprehensiveOrganic Synthesis, Volume 3, Chapters 2,4 and Sonogashira, Synthesis1977, 777. For further details of the Stille coupling reaction see: J.K. Stille, Angew. Chem. Int. Ed. 1986, 25, 508-524, M. Pereyre et al.,Tin in Organic Synthesis (Butterworths, Boston, 1987) pp 185-207 passim,and a review of synthetic applications in T. N. Mitchell, Synthesis1992, 803-815. The Buchwald reaction allows for the substitution ofamines, both primary and secondary, as well as 1 H-nitrogen heterocyclesat the aryl bromide. For further details of the Buchwald reaction see J.F. Hartwig, Angew. Chem. Int. Ed. 1998, 37, 2046-2067.

Scheme 4 illustrates the modification of the N-terminal and C-teminal ofthe macrocycle. Deprotection of the Boc moiety with an acid, such as,but not limited to hydrochloric acid yields compounds of formula (4-2).The amino moiety of formula (4-2) can be alkylated or acylated withappropriate alkyl halide or acyl groups to give compounds of formula(4-3). Compounds of formula (4-3) can be hydrolyzed with base such aslithium hydroxide to free up the acid moiety of formula (4-4).Subsequent activation of the acid moiety followed by treatment withappropriate acyl or sulfonyl groups to provide compounds of formula(4-5).

The sulfonamides (5-2) were prepared from the corresponding acids (5-1)by subjecting the acid to a coupling reagent (i.e. CDI, HATU, DCC, EDCand the like) at RT or at elevated temperature, with the subsequentaddition of the corresponding sulfonamide R₃—S(O)₂—NH₂ in the presenceof base wherein R₃ is as previously defined.

EXAMPLES

The compounds and processes of the present invention will be betterunderstood in connection with the following examples, which are intendedas an illustration only and not to limit the scope of the invention.Various changes and modifications to the disclosed embodiments will beapparent to those skilled in the art and such changes and modificationsincluding, without limitation, those relating to the chemicalstructures, substituents, derivatives, formulations and/or methods ofthe invention may be made without departing from the spirit of theinvention and the scope of the appended claims.

U.S. Patent Application Publication No. 20050153877 also describescertain compounds where G=OH.

Example 1 Synthesis of the Cyclic Peptide Precursor

1A. To a solution of Boc-L-2-amino-8-nonenoic acid 1a (1.36 g, 5 mol)and the commercially available cis-L-hydroxyproline methyl ester 1b(1.09 g, 6 mmol) in 15 ml DMF, was added DIEA (4 ml, 4eq.) and HATU (4g, 2eq). The coupling was carried out at 0° C. over a period of 1 hour.The reaction mixture was diluted with 100 mL EtOAc, and followed bywashing with 5% citric acid 2×20 ml, water 2×20 ml, 1M NaHCO₃ 4×20 mland brine 2×10 ml, respectively. The organic phase was dried overanhydrous Na₂SO₄ and then was evaporated, affording the dipeptide 1c(1.91g, 95.8%) that was identified by HPLC (Retention time=8.9 min,30-70%, 90% B), and MS (found 421.37, M+Na⁺).

1B. The dipeptide 1c (1.91 g) was dissolved in 15 mL of dioxane and 15mL of 1 N LiOH aqueous solution and the hydrolysis reaction was carriedout at RT for 4 hours. The reaction mixture was acidified by 5% citricacid and extracted with 100 mL EtOAc, and followed by washing with water2×20 ml, and brine 2×20 ml, respectively. The organic phase was driedover anhydrous Na₂SO₄ and then removed in vacuum, yielding the freecarboxylic acid compound 1d (1.79 g, 97%), which was used for next stepsynthesis without need for further purification.

1C. To a solution of the free acid obtained above (1.77, 4.64 mmol) in 5ml DMF, D-β-vinyl cyclopropane amino acid ethyl ester 1e (0.95 g, 5mmol), DIEA (4 ml, 4eq.) and HATU (4 g, 2eq) were added. The couplingwas carried out at 0° C. over a period of 5 hours. The reaction mixturewas diluted with 80 mL EtOAc, and followed by washing with 5% citricacid 2×20 ml, water 2×20 ml, IM NaHCO₃ 4×20 ml and brine 2×10 ml,respectively. The organic phase was dried over anhydrous Na₂SO₄ and thenevaporated. The residue was purified by silica gel flash chromatographyusing different ratios of hexanes:EtOAc as elution phase(5:1→3:1→1:1→1:2→1:5). The linear tripeptide 1f was isolated as an oilafter removal of the elution solvents (1.59 g, 65.4%), identified byHPLC (Retention time=11.43 min) and MS (found 544.84, M+Na⁺).

1D. Ring Closing Metathesis (RCM). A solution of the linear tripeptide1f (1.51 g, 2.89 mmol) in 200 ml dry DCM was deoxygenated by bubblingN₂. Hoveyda's 1^(st) generation catalyst (5 mol % eq.) was then added assolid. The reaction was refluxed under N₂ atmosphere 12 hours. Thesolvent was evaporated and the residue was purified by silica gel flashchromatography using different ratios of hexanes:EtOAc as elution phase(9:1→5:1→3:1→1:1→1:2→1:5). The cyclic peptide precursor 1 was isolatedas a white powder after removal of the elution solvents (1.24 g, 87%),identified by HPLC (Retention time=7.84 min, 30-70%, 90% B), and MS(found 516.28, M+Na⁺). For further details of the synthetic methodsemployed to produce the cyclic peptide precursor 1, see U.S. Pat. No.6,608,027, which is herein incorporated by reference in its entirety.

Example 2 Synthesis of the Cyclic Peptide Precursor Mesylate

2A. To a solution of the macrocyclic peptide precursor 1 (500 mg, 1.01mmol) and DIEA (0.4 ml, 2 mmol) in 2.0 ml DCM, mesylate chloride (0.1ml) was added slowly at 0° C. where the reaction was kept for 3 hours.30 mL EtOAc was then added and followed by washing with 5% citric acid2×10 ml, water 2×10 ml, IM NaHCO₃ 2×10 ml and brine 2×10 ml,respectively. The organic phase was dried over anhydrous Na₂SO₄ andevaporated, yielding the title compound mesylate that was used for nextstep synthesis without need for further purification.

Example 3 Compound of Formula IX wherein A=Boc,

Step 3a: Alkyne Formation

The alkyne of the current example,2-(2-thiazolyl)-4-methoxyphenylacetylene was prepared by adding to adegassed solution of 4 mmol of 4-ethynylanisole, 4 mmol of2-bromothiazole, and 1 ml of triethylamine in 10 ml of acetonitrile,140mg (0.2 mmol) of PdCl₂(PPh₃)₂ and 19 mg(0.1mmol) of CuI. The mixture wasdegassed and stirred for 5 minutes at RT and heated to 90° C. for 12hours. The reaction mixture was concentrated in vacuo and purified bysilica column to afford 0.61 g of brown liquid in a 70% yield.

MS (ESI): m/z=216.17 [M+H]

1HNMR (CDCl₃, 500 MHz) δ7.765(d, J=3 Hz, 1H), 7.472˜7.455(m, 2 H), 7.277(d, J=3.5 Hz, 1H), 6.837˜6.820 (m, 2H), 3.768 (s, 3H). Step 3b: TriazoleFormation

The 4-(2-thiazolyl)-5-(p-methoxyphenyl) triazole was prepared by addingto a pressure tube the compound (0.3 g) from step 3a, 0.74 ml oftrimethylsilyl azide, and 4 ml of xylenes and heating the mixture to140° C. for 48 hours. The reaction mixture was directly separated bysilica column to afford a brown liquid after purification (0.18 g, 50%).

MS (ESI): m/z=259.27 [M+H]

1HNMR (DMSO-d₆), 500 MHz) δ8.016(d, J=8.5 Hz, 2 H), 7.929(d, J=3 Hz, 1H), 7.817(d, J=3 Hz, 1 H), 7.066(d, J=8.5 Hz, 2H), 3.824(s, 3 H). Step3c

To a solution of 0.041 mmol of mesylate of macrocyclic precursor fromExample 2 and the compound (0.123 mmol) from step 3b in 1 ml of DMF wasadded 0.246 mmol cesium carbonate The reaction mixture was atirred at70° C. for 12 hours. The reaction mixture was extracted with EtOAc,washed with 1 M sodium bicarbonate (2×30 ml ) and water (2×30 ml ), andconcentrated in vacuo to obtain ethyl ester.

MS (ESI): m/z =734.34 [M+H]

Step 3d

The title compound was prepared by dissolving the title compound fromstep 3 c in 2 mL of dioxane and 1 mL of 1 N LiOH aqueous solution. Theresulting reaction mixture was stirred at RT for 8 hours. The pH of thereaction mixture was adjusted to 3 with citric acid; then the reactionmixture was extracted with EtOAc, and washed with brine and water. Theorganic solution was concentrated in vacuo for purification by HPLCafforded a yellow powder after lyophilization (10 mg, yield 34%).

MS (ESI): m/z=706.33 [M+H]

1HNMR (DMSO-d₆, 500 MHz) δ12.283 (s, broad, 1 H), 8.750 (s, broad, 1H),8.014 (d, J=9 Hz, 2H), 7.938 (d, J=3.5 Hz, 1H), 7.852 (d, J=3.5 Hz, 1H),6.997 (d, J=8 Hz, 2 H), 6.927 (d, J=7, 1H), 5.555 (s, broad, 1H), 5.499(m, 1H), 5.298 (t, J=18 Hz and 9 Hz, 4.643 (t, J=16 Hz and 8 Hz, 1H),4.558 (d, J=11.5 Hz, 1H), 4.125˜4.093 (m, 2H), 3.802 (s, 3H),2.890˜2.847 (m, 1H), 2.542˜2.497(m, 2H), 2.123˜2.106 (m, 1H), 1.806(s,broad, 1H), 1.701˜1.663(m, 1H), 1.519(s, broad, 1H), 1.460˜1.435(m, 1H),1.314˜1.074(m, 16H).Example 4 to Example 21 were made with different triazoles following thesimilar procedures described in Example 3.

Example 4 Compound of Formula IX wherein A=Boc,

MS (ESI): m/z=676.44 [M+H].

Example 5 Compound of Formula IX wherein A=Boc,

MS (ESI): m/z=595.28 [M+H].

Example 6 Compound of Formula IX wherein A=Boc,

MS(ESI): m/z=595.42 [M+H].

Example 7 Compound of Formula IX wherein A=Boc,

MS (ESI): m/z=690.42 [M+Na].

Example 8 Compound of Formula IX wherein A=Boc

MS (ESI): m/z=677.88, 679,89 [M+H].

Example 9 Compound of Formula IX wherein A=Boc,

MS (ESI): m/z=772.11 [M+Na].

Example 10 Compound of Formula IX wherein A=Boc,

MS (ESI): m/z=657.99 [M+Na].

Example 11 Compound of Formula IX wherein A=Boc,

MS (ESI): m/z=705.31 [M+H].

Example 12 Compound of Formula IX wherein A=Boc

MS (ESI): m/z=695.30 [M+H].

Example 13 Compound of Formula IX wherein A=Boc,

MS (ESI): m/z=677.25 [M+H].

Example 14 Compound of Formula IX wherein

MS (ESI): m/z=700.34 [M+H].

Example 15 Compound of Formula IX wherein

MS (ESI): m/z=716.32 [M+H].

Example 16 Compound of Formula IX wherein

MS (ESI): m/z=716.36 [M+H].

Example 17 Compound of Formula IX wherein

MS (ESI):m/z=715.39 [M+H].

Example 18 Compound of Formula IX wherein

MS (ESI): m/z=699.36 [M+H].

Example 19 Compound of Formula IX wherein

MS (ESI): m/z=703.40 [M+H].

Example 20 Compound of Formula IX wherein

MS (ESI): m/z=713.38 [M+H].

Example 21 Compound of Formula IX wherein

MS (ESI): m/z=700.36 [M+H].

Example 22 Compound of Formula IX wherein A=Boc,

To a solution of the compound (30 mg) from step 3d of Example 3 in DMFwas added CDI (10 mg). The reaction mixture was stirred at 40° C. for 1h and then added cyclopropylsulfonamide (11 mg) and DBU (12 μl). Thereaction mixture was stirred overnight at 40° C. The reaction mixturewas extracted with EtOAc. The organic extracts were washed with 1 MNaHCO₃, brine, dried over Na₂SO₄, filtered and concentrated. The residuewas purified by silica gel chromatograph to give desired product.

MS (ESI): m/z=809.30 [M+H].

Example 23 to Example 41 were made with different sulfonamides followingthe similar procedures described in Example 22.

Example 23 Compound of Formula IX wherein A=Boc,

MS (ESI): m/z=779.26 [M+H].

Example 24 Compound of Formula IX wherein A=Boc,

MS (ESI): m/z=698.36 [M+H].

Example 25 Compound of Formula IX wherein A=Boc,

MS (ESI): m/z=772.55 [M+H].

Example 26 Compound of Formula IX wherein A=Boc,

MS (ESI): m/z=880.51, 882.51 [M+H].

Example 27 Compound of Formula IX wherein A=Boc,

MS (ESI): m/z=852.60 [M+H].

Example 28 Compound of Formula IX wherein A=Boc,

MS (ESI): m/z=738.55 [M+H].

Example 29 Compound of formula IX wherein A=Boc,

MS (ESI): m/z=808.54 [M+H].

Example 30 Compound of Formula IX wherein A=Boc,

MS (ESI): m/z=798.57 [M+H].

Example 31 Compound of Formula IX wherein A=Boc,

MS (ESI): m/z=780.51 [M+H].

Example 32 Compound of Formula IX wherein

MS (ESI): m/z=803.47 [M+H].

Example 33 Compound of Formula IX wherein A=Boc,

MS (ESI): m/z=775.38 [M+H].

Example 34 Compound of Formula IX wherein A=Boc,

MS (ESI): m/z=812.34 [M+H].

Example 35 Compound of Formula IX wherein A=Boc,

MS (ESI):m/z=883.33, 885.31 [M+H].

Example 36 Compound of Formula IX wherein A=Boc,

MS (ESI): m/z=741.39 [M+H].

Example 37 Compound of Formula IX wherein

MS (ESI): m/z=806.30 [M+H].

Example 38 Compound of Formula IX wherein A=Boc,

MS (ESI): m/z=808.37 [M+H].

Example 39 Compound of Formula IX wherein A=Boc,

MS (ESI): m/z=845.33 [M+H].

Example 40 Compound of Formula IX wherein A=Boc.

MS (ESI): m/z=916.31, 918.31 [M+H].

Example 41 Compound of Formula IX wherein A=Boc,

MS (ESI): m/z=774.38 [M+H].

Example 42 Compound of Formula IX wherein

MS (ESI): m/z=839.28 [M+H].

Example 43 Compound of Formula IX wherein

Step 43a

The solution of the compound from Example 24 in 5 ml 4NHCl/Dioxne wasstirred at RT for 1 h. The reaction mixture was concentrated in vacuum.The residue was evaporated twice with DCM. The desired product wascarried out directly to the next step.

MS (ESI): m/z=598.26 [M+H].

Step 43b

To the solution of the compound from step 43a in 2 ml DCM was added DIEA(122 μl)) and cyclopentylchloroformate (0.216 mmol)). The reactionmixture was stirred at RT for 1 h. The reaction mixture was extractedwith EtOAc. The organic layer was washed with 1 M NaHCO₃, water, brine,dried over Na₂SO₄, filtered and concentrated. The residue was purifiedby HPLC to give the desired product.

MS (ESI): m/z=710.30 [M+H].

13 C(CD3OD): 177.9, 173.1, 169.4, 156.5, 143.9, 137.1, 135.4, 125.1,116.3, 77.5, 65.1, 60.4, 60.3, 53.6, 52.3, 43.9, 34.1, 32.6, 32.3, 32.2,32.1, 32.0, 30.7, 30.2, 27.3, 27.1, 26.4, 23.3, 23.2, 22.2, 21.3,19.7.

Example 44 to Example 92 (Formula IX) are made following the proceduresdescribed in Examples 22 or 42.

(IX)

Example # A Q G 44

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Example 93 Compound of formula IX wherein

Step 93a

The title compound was prepared using commercially availableBoc-cis-L-hydroxyproline and D-β-vinyl cyclopropane amine acid ethylester 1e via condition similar to those described in step 1c of Example1.

Step 93b

The title compound was prepared from Boc-protected dipeptide 93a viaconditions similar to those described in step 43a of Example 43.

Step 93c

The title compound was prepared using(S)-2-(cyclopentyloxycarbonyl)-amino-8-nonenoic acid (Organic ProcessResearch and Development (2007), 11(1), 60-63) and dipeptide 93b viaconditions similar to those described in step 1 a of Example 1.

MS (ESI): m/z=534.2 [M+H].

Step 93d

The title compound was prepared using diene 93c via conditions similarto those described in step ID of Example 1.

MS(ESI):m/z=506.2[M+H].

Step 93e

The title compound was prepared from macrocycle 93d and brosyl chloridevia conditions similar to those described in Example 2.

MS (ESI): m/z=712.2 [M+H].

Step 93f

The title compound was prepared from brosylated macrocycle 93e and4-bromo-5-phenyl-1,2,3-triazole (European J. of Med. Chem. (1983),18(5), 471) via conditions similar to those described in step 3c ofExample 3.

MS (ESI): m/z=711.1 [M+H].

Step 93 g

The title compound was prepared from ester 93f via conditions similar tothose described in step 3d of Example 3.

MS (ESI): m/z=685.5 [M+H].

Step 93h

A mixture of bromotriazole 93 g (50 mg, 0.073 mmol),quinolin-8-ylboronic acid (38.0 mg, 0.219 mmol), potassium phosphate(46.6 mg, 0.219 mmol),dicyclohexyl(2′,4′,6′-triisopropylbiphenyl-2-yl)phosphine (3.00 mg, 6.29μmol), and Pd₂(dba)₃ (0.980 mg, 1.070 μmol) was added to a microwavevial and diluted with acetonitrile (183 μl) and butanol (100 μL). Thevial was irradiated for 15min @ 140 ° C. The reaction mixture wasdiluted with 200 ul of 4N HCl in dioxane and the solvent removed under astream of nitrogen. The reaction mixture was purified by silica columnto afford a white solid after purification (40 mg 74%).

MS (ESI): m/z=711.1 [M+H].

Step 93

The title compound was prepared from acid 93 g via conditions similar tothose described in Example 22.

MS (ESI): m/z=835.1 [M+H].

Example 94 Compound of Formula IX wherein

Step 94a

The title compound was prepared from bromotriazole 93 g and2-fluoropyridin-3-ylboronic acid via conditions similar to thosedescribed in step 93h of Example 93.

MS (ESI): m/z=700.2 [M+H].

Step 94b

The title compound was prepared from acid 94a via conditions similar tothose described in Example 22.

MS (ESI): m/z=803.0 [M+H].

Example 95 Compound of formula IX wherein

Step 95a

The title compound was prepared from bromotriazole 93 g and2-fluoropyridin-4-ylboronic acid via conditions similar to thosedescribed in step 93h of Example 93.

MS (ESI): m/z=700.2 [M+H].

Step 95b

The title compound was prepared from acid 95a via conditions similar tothose described in Example 22

MS (ESI): m/z=803.1 [M+H].

Example 96 Compound of Formula IX wherein

Step 96a

The title compound was prepared from bromotriazole 93 g andphenylboronic acid via conditions similar to those described in step 93hof Example 93.

MS (ESI): m/z=681.7 [M+H].

Step 96b

The title compound was prepared from acid 96a via conditions similar tothose described in Example 22.

MS (ESI): m/z=784.2 [M+H].

Example 97 Compound of Formula IX wherein

Step 97a

The title compound was prepared from bromotriazole 93 g andthiophen-2-ylboronic acid via conditions similar to those described instep 93h of Example 93.

MS (ESI): m/z=687.2 [M+H].

Step 97b

The title compound was prepared from acid 97a via conditions similar tothose described in Example 22.

MS (ESI): m/z=790.7 [M+H].

Example 98 Compound of Formula IX wherein

Step 98a

The title compound was prepared from bromotriazole 93 g andnaphthalen-2-ylboronic acid via conditions similar to those described instep 93h of Example 93.

MS (ESI): m/z=731.2 [M+H].

Step 98b

The title compound was prepared from acid 98a via conditions similar tothose described in Example 22.

MS (ESI): m/z=834.2 [M+H].

Example 99 Compound of Formula IX wherein

Step 99a

The title compound was prepared from bromotriazole 93 g andthiophen-3-ylboronic acid via conditions similar to those described instep 93h of Example 93.

MS (ESI): m/z=687.2 [M+H].

Step 99b

The title compound was prepared from acid 99a via conditions similar tothose described in Example 22.

MS (ESI): m/z=790.1 [M+H].

Example 100 Compound of formula IX wherein

Step 100a

The title compound was prepared from bromotriazole 93 g and2-methoxypyrimidin-5-ylboronic acid via conditions similar to thosedescribed in step 93h of Example 93.

MS (ESI): m/z=713.6 [M+H].

Step 100b

The title compound was prepared from acid 100a via conditions similar tothose described in Example 22.

MS (ESI): m/z=816.1 [M+H].

Example 101 Compound of Formula IX wherein

Step 101a

The title compound was prepared from bromotriazole 93 g and4-methoxyphenylboronic acid via conditions similar to those described instep 93h of Example 93.

MS (ESI): m/z=712.0 [M+H].

Step 101b

The title compound was prepared from acid 101a via conditions similar tothose described in Example 22.

MS (ESI): m/z=814.0 [M+H].

Example 101 Compound of formula IX wherein

The title compound was prepared from bromotriazole 93 g via conditionssimilar to those described in Example 22.

MS (ESI): m/z=786.0 [M+H].

Example 102 Compound of formula IX wherein

A mixture of bromotriazole 101 (20 mg, 0.025 mmol),3-(4-fluorophenylcarbamoyl)phenylboronic acid (23.3 mg, 0.09 mmol),FibreCat 1007 (AlfaAesar, 21 mg, 0.0075 mmol), a 1 M solution ofpotassium carbonate (30 μL, 0.03 mmol) and ethanol (1 mL) was placed ina microwave tube and heat to 120° C. for 30 min under microwaveconditions. The reaction mixture was filtered through a Si-Carbonatecartridge (2 g, 0.79 mmol/g) and eluted with methanol. The solvent wasreduced and the residue purified by reverse phase HPLC to provide thetitle compound (3.5 mg, 15% yield).

MS (ESI): m/z=921.2 [M+H].

Example 103 Compound of formula IX wherein

The title compound was prepared from bromotriazole 101 and3-(furan-2-ylmethylcarbamoyl)phenylboronic acid via conditions similarto those described in Example 102.

MS (ESI): m/z=907.3 [M+H].

Example 104 Compound of Formula IX wherein

The title compound was prepared from bromotriazole 101 and4-(cyanomethyl)phenylboronic acid via conditions similar to thosedescribed in Example 102.

MS (ESI): m/z=823.6 [M+H].

Example 105 Compound of formula IX wherein

The title compound was prepared from bromotriazole 101 and6-methoxypyridin-3-ylboronic acid via conditions similar to thosedescribed in Example 102.

MS (ESI): m/z=815.2 [M+H].

Example 106 Compound of formula IX wherein

The title compound was prepared from bromotriazole 101 and6-(trifluoromethoxy)pyridin-3-ylboronic acid via conditions similar tothose described in Example 102.

MS (ESI): m/z=868.2 [M+H].

Example 107 Compound of formula IX wherein

The title compound was prepared from bromotriazole 101 and3-acetylphenylboronic acid via conditions similar to those described inExample 102.

MS (ESI): m/z=826.3 [M+H].

Example 108 Compound of formula IX wherein

The title compound was prepared from bromotriazole 101 and3-(morpholine-4-carbonyl)phenylboronic acid via conditions similar tothose described in Example 102.

MS (ESI): m/z=897.3 [M+H].

Example 109 Compound of formula IX wherein

The title compound was prepared from bromotriazole 101 and4-phenoxyphenylboronic acid via conditions similar to those described inExample 102.

MS (ESI): m/z=876.2 [M+H].

Example 110 Compound of formula IX wherein

Step 110a

The title compound was prepared from brosylated macrocycle 93e andcommercially available 4-(4-methoxyphenyl)-2H-1,2,3-triazole viaconditions similar to those described in step 3c of Example 3.

Step 110b

The title compound was prepared using the compound of step 110a viaconditions similar to those described in step 3d of Example 3.

MS (ESI): m/z=635.3 [M+H].

Step 110c

The title compound was prepared from acid 110b via conditions similar tothose described in Example 22.

MS (ESI): m/z=738.2 [M+H].

The compounds of the present invention exhibit potent inhibitoryproperties against the HCV NS3 protease. The following examples describeassays in which the compounds of the present invention can be tested foranti-HCV effects.

Example 111 NS3/NS4a Protease Enzyme Assay

HCV protease activity and inhibition is assayed using an internallyquenched fluorogenic substrate. A DABCYL and an EDANS group are attachedto opposite ends of a short peptide. Quenching of the EDANS fluorescenceby the DABCYL group is relieved upon proteolytic cleavage. Fluorescenceis measured with a Molecular Devices Fluoromax (or equivalent) using anexcitation wavelength of 355 nm and an emission wavelength of 485 nm.

The assay is run in Corning white half-area 96-well plates (VWR29444-312 [Corning 3693]) with full-length NS3 HCV protease 1b tetheredwith NS4A cofactor (final enzyme concentration 1 to 15 nM). The assaybuffer is complemented with 10 μM NS4A cofactor Pep 4A (Anaspec 25336 orin-house, MW 1424.8). RET S1(Ac-Asp-Glu-Asp(EDANS)-Glu-Glu-Abu-[COO]Ala-Ser-Lys-(DABCYL)-NH₂,-AnaSpec22991, MW 1548.6) is used as the fluorogenic peptide substrate. Theassay buffer contains 50 mM Hepes at pH 7.5, 30 mM NaCl and 10 mM BME.The enzyme reaction is followed over a 30 minutes time course at roomtemperature in the absence and presence of inhibitors.

The peptide inhibitors HCV Inh 1 (Anaspec 25345, MW 796.8)Ac-Asp-Glu-Met-Glu-Glu-Cys-OH, [−20° C.] and HCV Inh 2 (Anaspec 25346,MW 913.1) Ac-Asp-Glu-Dif-Cha-Cys-OH, are used as reference compounds.

IC50 values are calculated using XLFit in ActivityBase (IDBS) usingequation 205: y=A+((B−A)/1+((C/x)̂D)))

Example 112 Cell-Based Replicon Assay

Quantification of HCV replicon RNA (HCV Cell Based Assay) isaccomplished using the Huh 11-7 cell line (Lohmann, et al Science285:110-113, 1999). Cells are seeded at 4×10³ cells/well in 96 wellplates and fed media containing DMEM (high glucose), 10% fetal calfserum, penicillin-streptomycin and non-essential amino acids. Cells areincubated in a 7.5% CO₂ incubator at 37 ° C. At the end of theincubation period, total RNA is extracted and purified from cells usingAmbion RNAqueous 96 Kit (Catalog No. AM1812). To amplify the HCV RNA sothat sufficient material can be detected by an HCV specific probe(below), primers specific for HCV (below) mediate both the reversetranscription of the HCV RNA and the amplification of the cDNA bypolymerase chain reaction (PCR) using the TaqMan One-Step RT-PCR MasterMix Kit (Applied Biosystems catalog no. 4309169). The nucleotidesequences of the RT-PCR primers, which are located in the NS5B region ofthe HCV genome, are the following:

HCV Forward primer “RBNS5bfor” 5′GCTGCGGCCTGTCGAGCT: (SEQ ID NO: 1) HCVReverse primer “RBNS5Brev” 5′CAAGGTCGTCTCCGCATAC. (SEQ ID NO 2)

Detection of the RT-PCR product is accomplished using the AppliedBiosystems (ABI) Prism 7500 Sequence Detection System (SDS) that detectsthe fluorescence that is emitted when the probe, which is labeled with afluorescence reporter dye and a quencher dye, is degraded during the PCRreaction. The increase in the amount of fluorescence is measured duringeach cycle of PCR and reflects the increasing amount of RT-PCR product.Specifically, quantification is based on the threshold cycle, where theamplification plot crosses a defined fluorescence threshold. Comparisonof the threshold cycles of the sample with a known standard provides ahighly sensitive measure of relative template concentration in differentsamples (ABI User Bulletin #2 Dec. 11, 1997). The data is analyzed usingthe ABI SDS program version 1.7. The relative template concentration canbe converted to RNA copy numbers by employing a standard curve of HCVRNA standards with known copy number (ABI User Bulletin #2 Dec. 11,1997).

The RT-PCR product was detected using the following labeled probe:

(SEQ ID NO: 3) 5′ FAM-CGAAGCTCCAGGACTGCACGATGCT-TAMRA

-   -   FAM=Fluorescence reporter dye.    -   TAMRA:=Quencher dye.

The RT reaction is performed at 48 ° C. for 30 minutes followed by PCR.Thermal cycler parameters used for the PCR reaction on the ABI Prism7500 Sequence Detection System are: one cycle at 95 ° C., 10 minutesfollowed by 40 cycles each of which include one incubation at 95 ° C.for 15 seconds and a second incubation for 60 ° C. for 1 minute.

To normalize the data to an internal control molecule within thecellular RNA, RT-PCR is performed on the cellular messenger RNAglyceraldehyde-3-phosphate dehydrogenase (GAPDH). The GAPDH copy numberis very stable in the cell lines used. GAPDH RT-PCR is performed on thesame RNA sample from which the HCV copy number is determined. The GAPDHprimers and probesare contained in the ABI Pre-Developed TaqMan AssayKit (catalog no. 4310884E). The ratio of HCV/GAPDH RNA is used tocalculate the activity of compounds evaluated for inhibition of HCV RNAreplication.

Activity of Compounds as Inhibitors of HCV Replication (Cell BasedAssay) in Replicon Containing Huh-7 Cell Lines.

The effect of a specific anti-viral compound on HCV replicon RNA levelsin Huh-11-7cells is determined by comparing the amount of HCV RNAnormalized to GAPDH (e.g. the ratio of HCV/GAPDH) in the cells exposedto compound versus cells exposed to the DMSO vehicle (negative control).Specifically, cells are seeded at 4×10³ cells/well in a 96 well plateand are incubated either with: 1) media containing 1% DMSO (0%inhibition control), or 2) media/1% DMSO containing a fixedconcentration of compound. 96 well plates as described above are thenincubated at 37 ° C. for 4 days (EC50 determination). Percent inhibitionis defined as:

% Inhibition=100−100*S/C1

-   -   where    -   S=the ratio of HCV RNA copy number/GAPDH RNA copy number in the        sample;    -   C1=the ratio of HCV RNA copy number/GAPDH RNA copy number in the        0% inhibition control (media/1% DMSO).

The dose-response curve of the inhibitor is generated by adding compoundin serial, three-fold dilutions over three logs to wells starting withthe highest concentration of a specific compound at 1.5 μM and endingwith the lowest concentration of 0.23 nM. Further dilution series (500nM to 0.08 nM for example) is performed if the EC50 value is notpositioned well on the curve.

EC50 is determined with the IDBS Activity Base program “XL Fit” using a4-paramater, non-linear regression fit (model # 205 in version 4.2.1,build 16).

In the above assays, representative compounds of the present inventionare found to have HCV replication inhibitory activity and HCV NS3protease inhibitory activity. These compounds were also effective ininhibiting HCV NS3 proteases of different HCV genotypes includinggenotypes 1, 2, 3 and 4.

Representative compounds were tested in the above assays (Example 111and Example 112). The representative compounds disclosed here were foundto have activities in the ranges of ←0.2 nM-1000 nM in the NS3/NS4aProtease Enzyme Assay and ←0.2 nM-1000 nM in the Cell-Based RepliconAssay.

1. A compound of Formula I or II:

or a pharmaceutically acceptable salt, ester or prodrug thereof, whereinA is selected from —(C═O)—O—R₁, —(C═O)—R₂, —C(═O)—NH—R₂, and —S(O)₂—R₁,—S(O)₂NHR₂; R₁ is selected from the group consisting of: (i) aryl;substituted aryl; heteroaryl; substituted heteroaryl; (ii)heterocycloalkyl or substituted heterocycloalkyl; and (iii) —C₁-C₈alkyl, —C₂-C₈ alkenyl, or —C₂-C₈ alkynyl each containing 0, 1, 2, or 3heteroatoms selected from O, S, or N; substituted —C₁-C₈ alkyl,substituted —C₂-C₈ alkenyl, or substituted —C₂-C₈ alkynyl eachcontaining 0, 1, 2, or 3 heteroatoms selected from O, S or N; —C₃-C₁₂cycloalkyl, or substituted —C₃-C₁₂ cycloalkyl; —C₃-C₁₂ cycloalkenyl, orsubstituted —C₃-C₁₂ cycloalkenyl; R₂ is independently selected from thegroup consisting of: (i) hydrogen; (ii) aryl; substituted aryl;heteroaryl; substituted heteroaryl; (iii) heterocycloalkyl orsubstituted heterocycloalkyl; and (iv) —C₁-C₈ alkyl, —C₂-C₈ alkenyl, or—C₂-C₈ alkynyl each containing 0, 1, 2, or 3 heteroatoms selected fromO, S, or N; substituted —C₁-C₈ alkyl, substituted —C₂-C₈ alkenyl, orsubstituted —C₂-C₈ alkynyl each containing 0, 1, 2, or 3 heteroatomsselected from O, S or N; —C₃-C₁₂ cycloalkyl, or substituted —C₃-C₁₂cycloalkyl; —C₃-C₁₂ cycloalkenyl, or substituted —C₃-C₁₂ cycloalkenyl; Gis selected from —NHS(O)₂—R₃ and —NH(SO₂)NR4R₅; R₃ is selected from: (i)aryl; substituted aryl; heteroaryl; substituted heteroaryl (ii)heterocycloalkyl or substituted heterocycloalkyl; and (iii) —C₁-C₈alkyl, —C₂-C₈ alkenyl, or —C₂-C₈ alkynyl each containing 0, 1, 2, or 3heteroatoms selected from O, S or N, substituted —C₁-C₈ alkyl,substituted —C₂-C₈ alkenyl, or substituted —C₂-C₈ alkynyl eachcontaining 0, 1, 2, or 3 heteroatoms selected from O, S or N; —C₃-C₁₂cycloalkyl, or substituted —C₃-C₁₂ cycloalkyl; —C₃-C₁₂ cycloalkenyl, orsubstituted —C₃-C₁₂ cycloalkenyl; provided that R₃ is not CH₂Ph orCH₂CH₂Ph; R₄ and R₅ are independently selected from: (i) hydrogen; (ii)aryl; substituted aryl; heteroaryl; substituted heteroaryl; (iii)heterocycloalkyl or substituted heterocycloalkyl; and (iv) —C₁-C₈ alkyl,—C₂-C₈ alkenyl, or —C₂-C₈ alkynyl each containing 0, 1, 2, or 3heteroatoms selected from O, S, or N; substituted —C₁-C₈ alkyl,substituted —C₂-C₈ alkenyl, or substituted —C₂-C₈ alkynyl eachcontaining 0, 1, 2, or 3 heteroatoms selected from O, S or N; —C₃-C₁₂cycloalkyl, or substituted —C₃-C₁₂ cycloalkyl; —C₃-C₁₂ cycloalkenyl, orsubstituted —C₃-C₁₂ cycloalkenyl; L is selected from —CH₂—, —O—, —S— and—S(O)₂—; X and Y are independently selected from: (i) hydrogen; (ii)aryl; substituted aryl; heteroaryl; substituted heteroaryl; (iii)heterocycloalkyl or substituted heterocycloalkyl; (iv) —C₁-C₈ alkyl,—C₂-C₈ alkenyl, or —C₂-C₈ alkynyl each containing 0, 1, 2, or 3heteroatoms selected from O, S, or N; substituted —C₁-C₈ alkyl,substituted —C₂-C₈ alkenyl, or substituted —C₂-C₈ alkynyl eachcontaining 0, 1, 2, or 3 heteroatoms selected from O, S or N; —C₃-C₁₂cycloalkyl, or substituted —C₃-C₁₂ cycloalkyl; —C₃-C₁₂ cycloalkenyl, orsubstituted —C₃-C₁₂ cycloalkenyl; and (v) —W—R₆, where W is absent, orselected from —O—, —S—, —NH—, —N(Me)—, —C(O)NH—, and —C(O)N(Me)—; R₆ isselected from the group consisting of: (a) hydrogen; (b) aryl;substituted aryl; heteroaryl; substituted heteroaryl (c)heterocycloalkyl or substituted heterocycloalkyl; and (d) —C₁-C₈ alkyl,—C₂-C₈ alkenyl, or —C₂-C₈ alkynyl each containing 0, 1, 2, or 3heteroatoms selected from O, S or N; substituted —C₁-C₈ alkyl,substituted —C₂-C₈ alkenyl, or substituted —C₂-C₈ alkynyl eachcontaining 0, 1, 2, or 3 heteroatoms selected from O, S or N; —C₃-C₁₂cycloalkyl, or substituted —C₃-C₁₂ cycloalkyl; —C₃-C₁₂ cycloalkenyl, orsubstituted —C₃-C₁₂ cycloalkenyl; alternatively, X and Y taken togetherwith the carbon atoms to which they are attached to form a cyclic moietywhich selected from aryl, substituted aryl, heteroaryl, or substitutedheteroaryl;

denotes a carbon-carbon single or double bond. j=0, 1,2,3,or4; k=1 , 2,or 3; m=0, 1, or 2; and n=1,2or3.
 2. The compound of claim 1, whereinthe compound is of Formula III or IV:

or a pharmaceutically acceptable salt, ester or prodrug thereof, whereA, G, X and Y are as previously defined in claim
 1. 3. The compound ofclaim 1, wherein the compound is of Formula V or VI:

or a pharmaceutically acceptable salt, ester or prodrug thereof, whereX₁-X₄ are independently selected from —CR₇ and N, wherein R₇ isindependently selected from: (i) hydrogen; halogen; —NO₂; —CN; (ii)—M—R₄, M is O, S, NH, where R₄ is as previously defined in claim 1;(iii) NR₄R₅, where R₄ and R₅ are as previously defined in claim 1; (iv)—C₁-C₈ alkyl, —C₂-C₈ alkenyl, or —C₂-C₈ alkynyl each containing 0, 1, 2,or 3 heteroatoms selected from O, S, or N; substituted —C₁-C₈ alkyl,substituted —C₂-C₈ alkenyl, or substituted —C₂-C₈ alkynyl eachcontaining 0, 1, 2, or 3 heteroatoms selected from O, S or N; —C₃-C₁₂cycloalkyl, or substituted —C₃-C₁₂ cycloalkyl; —C₃-C₁₂ cycloalkenyl, orsubstituted —C₃-C₁₂ cycloalkenyl; (v) aryl; substituted aryl;heteroaryl; substituted heteroaryl; and (vi) heterocycloalkyl orsubstituted heterocycloalkyl; where A and G are as previously defined inclaim
 1. 4. The compound of claim 1, wherein the compound is of FormulaVII or VIII:

or a pharmaceutically acceptable salt, ester or prodrug thereof, whereY₁-Y₃ are independently selected from CR₇, N, NR₇, S and O; wherein R₇is independently selected from: (i) hydrogen; halogen; —NO₂; —CN; (ii)—M—R₄, M is O, S, NH, where R₄ is as previously defined in claim 1;(iii) NR₄R₅, where R₄ and R₅ are as previously defined in claim 1; (iv)—C₁-C₈ alkyl, —C₂-C₈ alkenyl, or —C₂-C₈ alkynyl each containing 0, 1, 2,or 3 heteroatoms selected from O, S, or N; substituted —C₁-C₈ alkyl,substituted —C₂-C₈ alkenyl, or substituted —C₂-C₈ alkynyl eachcontaining 0, 1, 2, or 3 heteroatoms selected from O, S or N; —C₃-C₁₂cycloalkyl, or substituted —C₃-C₁₂ cycloalkyl; —C₃-C₁₂ cycloalkenyl, orsubstituted —C₃-C₁₂ cycloalkenyl; (v) aryl; substituted aryl;heteroaryl; substituted heteroaryl; and (vi) heterocycloalkyl orsubstituted heterocycloalkyl; where A and G are as previously defined inclaim
 1. 5. A compound according to claim 1 which is selected fromcompounds of Formula IX, Table
 1. TABLE 1 (IX)

Example # A Q G 22

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109

110


6. A compound having a formula selected from formulae I, II, III, IV, V,VI, VII, VIII or IX as described in the specification, or apharmaceutically acceptable salt, ester or prodrug thereof
 7. Apharmaceutical composition comprising (1) a compound having a formulaselected from formulae I, II, III, IV, V, VI, VII, VIII or IX, asdescribed in the specification, or (2) a pharmaceutically acceptablesalt, ester or prodrug of said compound.
 8. A pharmaceutical compositioncomprising an inhibitory amount of a compound according to claim 1 or apharmaceutically acceptable salt, ester, or prodrug thereof, incombination with a pharmaceutically acceptable carrier or excipient. 9.A method of treating a hepatitis C viral infection in a subject,comprising administering to the subject an inhibitory amount of apharmaceutical composition according to claim
 8. 10. A method ofinhibiting the replication of hepatitis C virus, the method comprisingsupplying a hepatitis C viral NS3 protease inhibitory amount of thepharmaceutical composition of claim
 8. 11. The method of claim 9 furthercomprising administering concurrently an additional anti-hepatitis Cvirus agent.
 12. The method of claim 11, wherein said additionalanti-hepatitis C virus agent is selected from the group consisting of:α-interferon, β-interferon, ribavarin, and adamantine.
 13. The method ofclaim 11, wherein said additional anti-hepatitis C virus agent is aninhibitor of hepatitis C virus helicase, polymerase, metalloprotease, orIRES.
 14. A process of making a compound having a formula selected fromformulae I, II, III, IV, V, VI, VII, VIII or IX, as described in thespecification, according to the schemes and examples described therein.15. A pharmaceutical composition comprising a compound of claim 1, or apharmaceutically acceptable salt, ester, or prodrug thereof.
 16. Thepharmaceutical composition of claim 15, further comprising anotheranti-HCV agent.
 17. The pharmaceutical composition of claim 15, furthercomprising an agent selected from interferon, ribavirin, amantadine,another HCV protease inhibitor, an HCV polymerase inhibitor, an HCVhelicase inhibitor, or an internal ribosome entry site inhibitor. 18.The pharmaceutical composition of claim 15, further comprising pegylatedinterferon.
 19. The pharmaceutical composition of claim 15, furthercomprising another anti-viral, anti-bacterial, anti-fungal oranti-cancer agent, or an immune modulator.